JP2022543062A - Dosing Regimens for IDO Inhibitors - Google Patents

Abstract

The present disclosure relates to administration regimens for treating cancer by administering epacadostat in combination with an antibody or antibody fragment thereof that binds PD-1.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 62/881,518, filed Aug. 1, 2019, the disclosure of which is hereby incorporated by reference in its entirety. be

The present disclosure relates to administration regimens for treating cancer by administering epacadostat in combination with an antibody or antibody fragment thereof that binds PD-1.

Tryptophan (Trp) is an essential amino acid required for the biosynthesis of proteins, niacin and the neurotransmitter 5-hydroxytryptamine (serotonin). The enzyme indoleamine 2,3-dioxygenase (also known as INDO, IDO, or IDO1) catalyzes the first rate-limiting step in the breakdown of L-tryptophan to N-formyl-kynurenine. In human cells, Trp depletion due to IDO activity is a prominent gamma interferon (IFN-γ)-induced antimicrobial effector mechanism. IFN-γ stimulation induces activation of IDO, which leads to depletion of Trp, thereby blocking the growth of Trp-dependent intracellular pathogens such as Toxoplasma gondii and Chlamydia trachomatis. IDO activity also has antiproliferative effects on many tumor cells, and IDO induction was observed in vivo during allogeneic tumor rejection, indicating a possible role for this enzyme in the tumor rejection process (Daubener, et al. al., 1999, Adv. Exp. Med. Biol., 467:517-24, Taylor, et al., 1991, FASEB J., 5:2516-22).

HeLa cells co-cultured with peripheral blood lymphocytes (PBL) have been observed to acquire an immunosuppressive phenotype through upregulation of IDO activity. The decrease in PBL proliferation upon treatment with interleukin-2 (IL2) was thought to result from IDO released by tumor cells in response to IFNG secretion by PBLs. This effect was reversed by treatment with the specific IDO inhibitor, 1-methyl-tryptophan (1MT). It has been proposed that IDO activity in tumor cells may play a role in compromising anti-tumor responses (Logan, et al., 2002, Immunology, 105:478-87).

Recently, the immunomodulatory role of Trp depletion has received much attention. Several lines of evidence suggest that IDO is involved in the induction of immune tolerance. Studies of mammalian pregnancy, tumor resistance, chronic infections and autoimmune diseases have shown that cells expressing IDO can suppress T cell responses and promote tolerance. For example, increased levels of IFN and elevated levels of urinary Trp metabolites have been observed in autoimmune diseases, and the systemic or local depletion of Trp that occurs in autoimmune diseases contributes to the degeneration and It has been hypothesized that it may be associated with debilitating symptoms.

Further evidence for a tumor immune resistance mechanism based on tryptophan degradation by IDO suggests that most human tumors constitutively express IDO, and that expression of IDO by immunogenic mouse tumor cells reduces their expression by pre-immunized mice. It follows from the observation that it prevents rejection. This effect is accompanied by a lack of accumulation of specific T cells at the tumor site and can be partially reversed by systemic treatment of mice with inhibitors of IDO in the absence of significant toxicity. Therefore, it was suggested that the efficacy of therapeutic vaccination of cancer patients could be improved by concomitant administration of IDO inhibitors (Uyttenhove et al., 2003, Nature Med., 9:1269-74). An IDO inhibitor, 1-MT, has also been shown to be able to synergize with chemotherapeutic agents to reduce tumor growth in mice, and IDO inhibition also enhances the antitumor activity of conventional cytotoxic therapies. (Muller et al., 2005, Nature Med., 11:312-9).

One mechanism contributing to immunological unresponsiveness to tumors may be the presentation of tumor antigens by tolerogenic host APCs. A subset of human IDO-expressing antigen-presenting cells (APCs) that co-expressed CD123 (IL3RA) and CCR6 and inhibited T-cell proliferation has also been described. Both mature and immature CD123-positive dendritic cells suppressed T cell activity, and this IDO suppressive activity was blocked by 1MT (Munn, et al., 2002, Science, 297:1867-70). It has also been demonstrated that mouse tumor-draining lymph nodes (TDLNs) contain a subset of plasmacytoid dendritic cells (pDCs) that constitutively express immunosuppressive levels of IDO. In vitro, despite containing only 0.5% lymph node cells, these pDCs potently suppressed T-cell responses to antigens presented by the pDCs themselves and, in a dominant manner, non-suppressed suppressed T cell responses to third party antigens presented by sexual APCs. Within the pDC population, the majority of functional IDO-mediated suppressor activity segregated with a novel pDC subset co-expressing the B-lineage marker CD19. Therefore, it was hypothesized that IDO-mediated suppression by pDCs in TDLNs creates a local microenvironment that potently suppresses host anti-tumor T cell responses (Munn, et al., 2004, J. Clin. Invest., 114 ( 2): 280-90).

IDO breaks down the indole portions of tryptophan, serotonin and melatonin, initiating the production of neuroactive and immunomodulatory metabolites collectively known as kynurenines. By locally depleting tryptophan and increasing proapoptotic kynurenines, IDO expressed by dendritic cells (DCs) can profoundly affect T cell proliferation and survival. IDO induction in DCs may be a common mechanism of regulatory T cell-driven deletional tolerance. Since such tolerogenic responses can be expected to operate under a variety of physiopathological conditions, tryptophan metabolism and kynurenine production may represent an important interface between the immune and nervous systems (see Grohmann, et al., 2003, Trends Immunol., 24:242-8). In conditions of persistent immune activation, the availability of free serum Trp is reduced and serotonergic function may also be affected as a result of decreased serotonin production (Wirleitner, et al., 2003, Curr Chem., 10:1581-91).

In light of experimental data demonstrating the role of IDO in immunosuppression and tumor resistance and/or rejection, therapeutic agents aimed at inhibiting tryptophan degradation by inhibiting IDO activity are desirable. One potent inhibitor of IDO1 is epacadostat (INCB24360; 4-({2-[(aminosulfonyl)amino]ethyl}amino)-N-(3-bromo-4-fluorophenyl)- having the formula N′-Hydroxy-1,2,5-oxadiazole-3-carboximidamide):

There remains a need for new therapeutic regimens against cancer that use IDO1 inhibitors. The present disclosure is directed to this need and other needs.

The present disclosure provides, inter alia, a method of treating cancer in a patient comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 400 mg to about 700 mg on a free base basis twice daily;
(ii) an antibody, or antigen-binding fragment thereof, that binds to human PD-1, wherein the antibody comprises (ii-1) variable heavy (VH) complementarity determining regions (CDR) 1, VH CDR2, and VH CDR3; administering an antibody, or antigen-binding fragment thereof, comprising a VH domain and (ii-2) a VL domain comprising variable light (VL) CDR1, VL CDR2, and VL CDR3;
(a) the VH CDR1 comprises the amino acid sequence SYWMN (SEQ ID NO: 6);
(b) VH CDR2 comprises the amino acid sequence VIHPSDSETWLDQKFKD (SEQ ID NO: 7);
(c) the VH CDR3 comprises the amino acid sequence EHYGTSPFAY (SEQ ID NO: 8);
(d) the VL CDR1 comprises the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 9);
(e) VL CDR2 comprises the amino acid sequence AASNQGS (SEQ ID NO: 10);
(f) providing the method, wherein the VL CDR3 comprises the amino acid sequence QQSKEVPYT (SEQ ID NO: 11);

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 600 mg on a free base basis twice daily.

The disclosure further provides a method of treating cancer in a patient, comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 400 mg to about 700 mg twice daily on a free base basis; and (ii) an antibody that binds human PD-1, which is Antibody X. including administering.

Antibody X is retifanlimab. Surprisingly, doses of epacadostat (e.g., 600 mg) in the methods of the present disclosure, when administered in combination with Antibody X, compared to lower doses (e.g., 100 mg twice daily) reduced kynurenine levels to (see Example 1 below). While not wishing to be bound by any particular theory, it is believed that the claimed doses of epacadostat function by blocking additional IDO1 activity induced as a result of immune system stimulants such as Antibody X. be done.

ヒトＰＤ－１タンパク質（Ｇｅｎｂａｎｋ受託番号ＮＰ＿００５００９）のアミノ酸配列は、ＭＱＩＰＱＡＰＷＰＶＶＷＡＶＬＱＬＧＷＲＰＧＷＦＬＤＳＰＤＲＰＷＮＰＰＴＦＳＰＡＬＬＶＶＴＥＧＤＮＡＴＦＴＣＳＦＳＮＴＳＥＳＦＶＬＮＷＹＲＭＳＰＳＮＱＴＤＫＬＡＡＦＰＥＤＲＳＱＰＧＱＤＣＲＦＲＶＴＱＬＰＮＧＲＤＦＨＭＳＶＶＲＡＲＲＮＤＳＧＴＹＬＣＧＡＩＳＬＡＰＫＡＱＩＫＥＳＬＲＡＥＬＲＶＴＥＲＲＡＥＶＰＴＡＨＰＳＰＳＰＲＰＡＧＱＦＱＴＬＶＶＧＶＶＧＧＬＬＧＳＬＶＬＬＶＷＶＬＡＶＩＣＳＲＡＡＲＧＴＩＧＡＲＲＴＧＱＰＬＫＥＤＰＳＡＶＰＶＦＳＶＤＹＧＥＬＤＦＱＷＲＥＫＴＰＥＰＰＶＰＣＶＰＥＱＴＥＹＡＴＩＶＦＰＳＧＭＧＴＳＳＰＡＲＲＧＳＡＤＧＰＲＳＡＱＰＬＲＰＥＤＧＨＣＳＷＰＬ（配列番号１）である。

Antibody X is a humanized IgG4 monoclonal antibody that binds to human PD-1 (see WO2017019846, which is incorporated herein by reference in its entirety). The amino acid sequences of the mature antibody X heavy and light chains are given below.

Complementarity determining regions (CDRs) 1, 2, and 3 of the variable heavy (VH) and variable light (VL) domains are shown in order from N-terminus to C-terminus of the mature VL and VH sequences, both underlined. and in bold. An antibody consisting of a mature heavy chain (SEQ ID NO:2) and a mature light chain (SEQ ID NO:3) listed below is referred to as Antibody X.

Mature antibody X heavy chain (HC)

Mature antibody X light chain (LC)

The variable heavy (VH) domain of Antibody X has the following amino acid sequence.

The variable light (VL) domain of Antibody X has the following amino acid sequence.

The amino acid sequences of the VH CDRs of Antibody X are listed below.
VH CDR1: SYWMN (SEQ ID NO: 6),
VH CDR2: VIHPSDSETWLDQKFKD (SEQ ID NO: 7),
VH CDR3: EHYGTSPFAY (SEQ ID NO: 8)

The amino acid sequences of the VL CDRs of Antibody X are listed below:
VL CDR1: RASESVDNYGMSFMNW (SEQ ID NO: 9),
VL CDR2: AASNQGS (SEQ ID NO: 10), and VL CDR3: QQSKEVPYT (SEQ ID NO: 11).

Accordingly, the present disclosure provides a method of treating cancer in a patient comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 400 mg to about 700 mg on a free base basis twice daily;
(ii) an antibody that binds to human PD-1, wherein the antibody (ii-1) a VH domain comprising a variable heavy (VH) complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3; -2) administering an antibody comprising a VL domain comprising variable light (VL) CDR1, VL CDR2, and VL CDR3;
(a) the VH CDR1 comprises the amino acid sequence SYWMN (SEQ ID NO: 6);
(b) VH CDR2 comprises the amino acid sequence VIHPSDSETWLDQKFKD (SEQ ID NO: 7);
(c) the VH CDR3 comprises the amino acid sequence EHYGTSPFAY (SEQ ID NO: 8);
(d) the VL CDR1 comprises the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 9);
(e) VL CDR2 comprises the amino acid sequence AASNQGS (SEQ ID NO: 10);
(f) providing the method, wherein the VL CDR3 comprises the amino acid sequence QQSKEVPYT (SEQ ID NO: 11);

In some embodiments, the antibody comprises an Fc region, wherein the Fc region is of IgG4 isotype. In some embodiments, the antibody comprises an Fc region of the IgG4 isotype and an IgG4 hinge domain comprising stabilizing mutations. In some embodiments, the antibody comprises an Fc region of the IgG4 isotype and an IgG4 hinge domain comprising the S228P substitution to reduce the occurrence of strand exchange (e.g., SEQ ID NO: 13: ESKYGPPCPPCP (Lu et al, (2008) ) "The Effect Of A Point Mutation On The Stability Of IgG4 As Monitored By Analytical Ultracentrifugation" J. Pharmaceutical Sciences 97:960-969)).

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof and Antibody X are administered to the patient simultaneously or sequentially. In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof and Antibody X are administered to the patient concurrently. In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof and Antibody X are administered to the patient sequentially.

In some embodiments the cancer is a solid tumor.

In some embodiments, the VH domain comprises the amino acid sequence shown in SEQ ID NO:4.

In some embodiments, the antibody comprises a heavy chain, and the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2.

In some embodiments, the VL domain comprises the amino acid sequence shown in SEQ ID NO:5.

In some embodiments, the antibody comprises a light chain, wherein the light chain comprises the amino acid sequence set forth in SEQ ID NO:3

In some embodiments, the VH domain comprises the amino acid sequence shown in SEQ ID NO:4 and the VL domain comprises the amino acid sequence shown in SEQ ID NO:5.

In some embodiments, the antibody comprises a heavy chain and a light chain, the heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2 and the light chain comprising the amino acid sequence set forth in SEQ ID NO:3.

In some embodiments, the antibody is a humanized antibody.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 500 mg to about 700 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 400 mg to about 600 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 500 mg to about 600 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 400 mg to about 600 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 550 mg to about 650 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 575 mg to about 625 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 400 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 425 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 450 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 475 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 500 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 525 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 550 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 575 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 600 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 625 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 650 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 675 mg on a free base basis twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered at a dose of about 700 mg on a free base basis twice daily.

In some embodiments, epacadostat is administered as the free base.

In some embodiments, epacadostat is administered at a dose of about 400 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 425 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 450 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 475 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 500 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 525 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 550 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 575 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 600 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 625 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 650 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 675 mg twice daily.

In some embodiments, epacadostat is administered at a dose of about 700 mg twice daily.

In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered as a pharmaceutical composition. In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered orally. In some embodiments, epacadostat or a pharmaceutically acceptable salt thereof is administered as a solid oral dosage form. In some embodiments, the solid oral dosage form is a tablet or capsule. In some embodiments, the solid oral dosage form is a tablet. In some embodiments, multiple tablets are administered to achieve the desired dose.

An anti-PD-1 antibody or antigen-binding fragment thereof can be administered to a subject, eg, a subject in need thereof, eg, a human subject, by a variety of methods. In many applications, the route of administration is one of intravenous injection or infusion (IV), subcutaneous injection (SC), intraperitoneal (IP), or intramuscular injection. Intra-articular delivery can also be used. Other methods of parenteral administration can also be used. Examples of such modalities include intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and epidural. and intrasternal injection. In some cases, administration can be oral.

The route and/or mode of administration of the antibody or antigen-binding fragment thereof may also be individualized, e.g., by monitoring the subject, e.g., to visualize tumors, e.g., using tomography. can be adjusted.

Antibodies or antigen-binding fragments can be administered as a fixed dose or in mg/kg patient body weight doses. Dosages can also be selected to reduce or avoid the production of antibodies to the antibody or antigen-binding fragment. Dosage regimens are adjusted to provide the desired response (eg, therapeutic response) or combination therapeutic effect. In general, a dose of antibody or antigen-binding fragment (and optionally a second agent) can be used to provide the subject with a bioavailable amount of the agent. For example, about 0.1-100 mg/kg, about 0.5-100 mg/kg, about 1 mg/kg-100 mg/kg, about 0.5-20 mg/kg, about 0.1-10 mg/kg, or about 1 Doses in the range of -10 mg/kg can be administered. Other doses can also be used. In certain embodiments, the antibody or antigen-binding fragment is administered to a subject in need of treatment at about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg, about 10 mg/kg. , about 15 mg/kg, about 20 mg/kg, about 30 mg/kg, about 35 mg/kg, or about 40 mg/kg. With respect to doses or dosages, the term "about" is ±10% of the recited dose, e.g., a dose of about 3 mg/kg is between 2.7 mg/kg patient weight and 3.3 mg/kg patient weight. is intended to indicate a range of

The composition comprises about 1 mg/mL to 100 mg/mL, or about 10 mg/mL to 100 mg/mL, or about 50-250 mg/mL, or about 100-150 mg/mL, or about 100-250 mg/mL of antibody or antigen It may contain a binding fragment.

Dosage unit form or "fixed dose" or "flat dose" as used herein refers to physically discrete units suited as unitary dosages for the treatment of subjects; each unit comprising the required It contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the pharmaceutical carrier and, optionally, other agents. Single or multiple doses may be administered. Alternatively, or additionally, antibodies or antigen-binding fragments thereof may be administered via continuous infusion. Exemplary fixed doses include about 375 mg, about 500 mg, and about 750 mg. With respect to doses or dosages, the term "about" is intended to indicate a range of ±10% of the recited dose, eg, a dose of about 375 mg goes from 337.5 mg to 412.5 mg.

Antibody or antigen-binding fragment doses can be, for example, at least 2 doses, 3 doses, 5 doses, 10 doses, or more, such as once or twice daily, or about 1-4 times, or preferably once a week, every other week (every 2 weeks), every 3 weeks, once a month, for example about 1-12 weeks, preferably 2-8 weeks, more preferably about 3-7 weeks , and even more preferably at periodic intervals over a period of time (course of treatment) sufficient to encompass about 4, 5, or 6 weeks. Factors that can influence the dosage and timing required to effectively treat a subject include, for example, the severity of the disease or disorder, the formulation, the route of delivery, previous treatments, the general health of the subject. and/or age, and other diseases present. Furthermore, treatment of a subject with a therapeutically effective amount of a compound can comprise a single treatment or, preferably, a series of treatments.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a fixed dose of about 375 mg once every three weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a fixed dose of 500 mg once every four weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a fixed dose of about 750 mg once every four weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 1 mg/kg once every two weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 3 mg/kg once every two weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 3 mg/kg once every four weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 10 mg/kg once every two weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 10 mg/kg once every four weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a fixed dose of about 375 mg once every three weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 500 mg once every four weeks.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment is administered at a dose of about 750 mg once every four weeks.

In some embodiments, the term "about" refers to ±10% of the value. Those skilled in the art will know that the values presented herein may vary due to experimental conditions such as data collection or instrument variability.

Epacadostat Epacadostat can be synthesized as described in US Pat. Nos. 8,088,803 and 9,321,755, which are incorporated herein by reference in their entireties.

The present disclosure also includes pharmaceutically acceptable salts of epacadostat described herein.

In some embodiments, epacadostat and salts thereof are substantially isolated. By "substantially isolated" is meant that the compound is at least partially or substantially separated from the environment in which epacadostat and its salts are formed or detected. A partial separation can include, for example, a concentrated composition of epacadostat. Substantial separation is defined as at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about Compositions containing 97% by weight, or at least about 99% by weight can be included. Methods of isolating compounds and their salts are routine in the art.

Epacadostat can exist in various solid forms. As used herein, "solid form" includes, for example, one or more of melting point, solubility, stability, crystallinity, hygroscopicity, water content, TGA characteristics, DSC characteristics, DVS characteristics, XRPD characteristics, etc. is meant to refer to a solid body characterized by the properties of Solid forms can be, for example, amorphous, crystalline, or mixtures thereof.

Different crystalline solid forms typically have different crystal lattices (eg, unit cells) and, as a result, usually different physical properties. In some cases, different crystalline solid forms have different water or solvent contents. Different crystal lattices can be identified by solid-state characterization methods, for example, by X-ray powder diffraction (XRPD). Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), etc., further aid in characterizing solid forms, as well as stability and solvent/water content. Helps determine quantity.

In some embodiments, the solid form is a crystalline form. In some embodiments, epacadostat is a crystalline solid described in US Pat. No. 8,088,803. In some embodiments, the solid form is substantially anhydrous (e.g., less than about 1% water, less than about 0.5% water, less than about 1.5% water, less than about 2% containing water). For example, water content is determined by Karl Fischer titration. In some embodiments, the solid form is characterized by a melting point, or DSC endotherm, centered at about 162 to about 166°C. In some embodiments, the solid form is characterized by a melting point or DSC endotherm centered at about 164°C. In some embodiments, the solid form has a weight loss of 0.3% when heated from 20°C to 150°C at a heating rate of 10°C/min.

In further embodiments, the solid form is selected from about 18.4°, about 18.9°, about 21.8°, about 23.9°, about 29.2°, and about 38.7° 2-theta. have at least one, two, or three XRPD peaks.

In some embodiments, the crystalline form has one or more of the peaks from the list of 2-theta peaks provided in the table below.

An XRPD pattern of reflections (peaks) is typically taken as a fingerprint of a particular crystalline form. It is well known that the relative intensities of XRPD peaks can vary widely depending, among other things, on the sample preparation technique, the crystal size distribution, the various filters used, the sample mounting procedure, and the particular instrument used. . In some cases, new peaks may be observed or existing peaks may disappear, depending on the instrument type or settings. As used herein, the term "peak" refers to reflections having a relative height/intensity of at least about 4% of the maximum peak height/intensity. In addition, instrument variation and other factors can affect the 2-theta values. Thus, peak assignments such as those reported herein may vary by plus or minus about 0.2° (2θ), and the term "substantially" is used herein in the context of XRPD. If so, it is meant to include the above changes.

Similarly, temperature readings for DSC, TGA, or other thermal experiments can vary by about ±3°C depending on instrumentation, specific settings, sample preparation, and the like.

A pharmaceutical composition can include a “therapeutically effective amount” of an agent described herein. Such effective amounts can be determined based on the effect of the administered agent, or the combined effect of agents when more than one agent is used. A "therapeutically effective amount" of an agent also includes factors such as the individual's disease state, age, sex, and weight, as well as the ability of the compound to elicit a desired response in an individual, e.g., amelioration of at least one disorder parameter, or at least one may vary due to amelioration of symptoms of one disorder. A therapeutically effective amount is also one in which any toxic or detrimental effects of the composition are outweighed by the therapeutically beneficial effects.

Preparation of Antibodies and Pharmaceutical Compositions of Antibodies In certain embodiments, an antibody that binds to human PD-1 comprises human heavy and light chain constant regions. In certain embodiments, the heavy chain constant region comprises the CH1 domain and hinge region. In some embodiments, the heavy chain constant region comprises a CH3 domain. If the heavy chain constant region contains substitutions, such substitutions modify a property of the antibody (e.g., Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, or one of complement functions). increase or decrease one or more). In certain embodiments, the antibody is an IgG antibody. In certain embodiments, the antibody is selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.

Antibodies, such as Antibody X, are prepared, for example, by preparing and expressing synthetic genes encoding the recited amino acid sequences, or by mutating human germline genes to provide genes encoding the recited amino acid sequences. can be made by Additionally, this antibody and other antibodies that bind to human PD-1 can be obtained using, for example, one or more of the following methods.

Humanized antibodies can be generated by replacing sequences of the Fv variable region that are not directly involved in antigen binding with equivalent sequences from human Fv variable regions. General methods for generating humanized antibodies are described by Morrison, S.; L. , Science, 229:1202-1207 (1985); Oi et al. , BioTechniques, 4:214 (1986), and US 5,585,089, US 5,693,761, US 5,693,762, US 5,859,205, and US 6,407,213. These methods involve isolating, manipulating, and expressing nucleic acid sequences encoding all or part of immunoglobulin Fv variable regions from at least one of a heavy or light chain. Sources of such nucleic acids are well known to those of skill in the art and can be obtained, for example, from hybridomas producing antibodies against a given target, from germline immunoglobulin genes, or from synthetic constructs, as described above. Recombinant DNA encoding the humanized antibody can then be cloned into an appropriate expression vector.

Human germline sequences are described, for example, in Tomlinson, I.; A. et al. , J. Mol. Biol. , 227:776-798 (1992); Cook, G.; P. et al. , Immunol. Today, 16:237-242 (1995); Chothia, D.; et al. , J. Mol. Bio. 227:799-817 (1992), and Tomlinson et al. , EMBOJ. , 14:4628-4638 (1995). The V BASE directory provides a comprehensive directory of human immunoglobulin variable region sequences (compiled by Tomlinson, IA et al., MRC Center for Protein Engineering, Cambridge, UK). These sequences can be used, for example, as a source of human sequence for framework regions and CDRs. Consensus human framework regions can also be used, for example, as described in US Pat. No. 6,300,064.

Other methods for humanizing antibodies can also be used. For example, other methods can account for the three-dimensional structure of antibodies, framework positions that are three-dimensionally proximal to binding determinants, and immunogenic peptide sequences. For example, WO 90/07861, U.S. Pat. 213, Tempest et al. (1991) Biotechnology 9:266-271. Yet another method is called "humaneering" and is described, for example, in US2005-008625.

The antibody can include a human Fc region, eg, a wild-type Fc region or an Fc region containing one or more modifications. In one embodiment, the constant region is altered (e.g., mutated to modify a property of the antibody) (e.g., Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, or increase or decrease one or more of the complement functions). For example, a human IgG1 constant region can be mutated at one or more residues, eg, one or more of residues 234 and 237 (based on Kabat numbering). Antibodies can have mutations in the CH2 region of the heavy chain that reduce or alter effector functions, such as Fc receptor binding and complement activation. For example, antibodies can have mutations as described in US Pat. Nos. 5,624,821 and 5,648,260. Antibodies may also be modified from immunoglobulins, such as mutations in the hinge region of IgG4 (eg, Angal et al. (1993) Mol. Immunol. 30:105-08), as disclosed in the art. Mutations that stabilize disulfide bonds between heavy chains may also be present. For example, U.S.A. S. See also 2005/0037000.

Antibodies that bind human PD-1 or human PD-L1 can be in the form of full-length antibodies or human PD-1 or human PD-L1, eg, Fab, Fab′, F(ab′) ₂ , Fv , Fd, dAbs, scFv, and low molecular weight forms of antibodies (eg, biologically active antibody fragments or minibodies) that bind to sc(Fv)2. Other antibodies encompassed by this disclosure include single domain antibodies (sdAb) containing a single variable chain, such as VH or VL, or biologically active fragments thereof. For example, Moller et al. , J. Biol. Chem. , 285(49):38348-38361 (2010), Harmsen et al. , Appl. Microbiol. Biotechnol. , 77(1):13-22 (2007), US 2005/0079574 and Davies et al. (1996) Protein Eng. , 9(6):531-7. Like whole antibodies, sdAbs can selectively bind to specific antigens. With a molecular weight of only 12-15 kDa, sdAbs are much smaller than common antibodies and even smaller than Fab fragments and single-chain variable fragments.

Provided herein are compositions comprising an antibody, or antigen-binding fragment thereof, that binds to human PD-1 or human PD-L1, and a mixture of one or more acidic variants, e.g. is less than about 80%, 70%, 60%, 60%, 50%, 40%, 30%, 30%, 20%, 10%, 5%, or 1%. Also provided is a composition comprising an antibody, or antigen-binding fragment thereof, that binds to human PD-1 or human PD-L1, comprising at least one deamidation site, wherein the pH of the composition is, for example, at least about 90% From about 5.0 to about 6.5 so that the antibody is not deamidated (ie, less than about 10% of the antibody is deamidated). In certain embodiments, less than about 5%, 3%, 2%, or 1% of the antibody is deamidated. The pH may be 5.0-6.0, such as 5.5 or 6.0. In certain embodiments, the pH of the composition is 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.5. 4, or 6.5.

An "acidic variant" is a variant of a polypeptide of interest that is more acidic (eg, as determined by cation exchange chromatography) than the polypeptide of interest. Examples of acidic mutants are deamidating mutants.

A "deamidated" variant of a polypeptide molecule has one or more asparagine residue(s) of the original polypeptide converted to aspartic acid, i.e., the neutral amide side chains are reduced to an overall acidic Polypeptides that have been converted to residues that have properties.

As used herein, the term "mixture" refers to a composition comprising an antibody or antigen-binding fragment thereof that binds to human PD-1 or human PD-L1. The presence of both the desired antibody, or antigen-binding fragment thereof, and one or more acidic variants thereof is implied. Acidic variants may comprise predominantly deamidated antibodies that bind human PD-1 or human PD-L1, along with minor amounts of other acidic variant(s).

In certain embodiments, the binding affinity (K _D ), on rate (K _D on) and/or off rate (K _D off) of an antibody that has been mutated to eliminate deamidation is, for example, about 5-fold , 2-fold, 1-fold (100%), 50%, 30%, 20%, 10%, 5%, 3%, 2% or less than 1% difference than that of the wild-type antibody.

Antibody Fragments Antibody fragments (eg, Fab, Fab', F(ab')2, Facb, and Fv) can be prepared by proteolysis of intact antibodies. For example, antibody fragments can be obtained by treating whole antibodies with enzymes such as papain, pepsin, or plasmin. Papain digestion of whole antibodies produces F(ab)2 or Fab fragments, pepsin digestion of whole antibodies produces F(ab')2 or Fab', and plasmin digestion of whole antibodies produces Facb fragments. do.

Alternatively, antibody fragments may be produced recombinantly. For example, a nucleic acid encoding an antibody fragment of interest can be constructed, introduced into an expression vector, and expressed in a suitable host cell. For example, Co, M.; S. et al. , J. Immunol. , 152:2968-2976 (1994); Better, M.; and Horwitz, A.; H. , Methods in Enzymology, 178:476-496 (1989); and Skerra, A.; , Methods in Enzymology, 178:476-496 (1989); Lamoyi, E.; , Methods in Enzymology, 121:652-663 (1989); Rousseaux, J.; et al. , Methods in Enzymology, (1989) 121:663-669 (1989), and Bird, R.; E. et al. , TIBTECH, 9:132-137 (1991)). All antibody fragments are E. These fragments can be easily produced in large amounts because they can be expressed in and secreted from E. coli. Antibody fragments can be isolated from antibody phage libraries. Alternatively, the Fab'-SH fragment can be obtained from E. It can be directly recovered from E. coli or chemically coupled to form F(ab)2 fragments (Carter et al., Bio/Technology 10:163-167 (1992)). According to another approach, F(ab')2 fragments can be isolated directly from recombinant host cell culture. Fab and F(ab')2 fragments containing salvage receptor binding epitope residues and having increased in vivo half-lives are described in US Pat. No. 5,869,046.

Minibodies Minibodies of antibodies that bind human PD-1 or human PD-L1 include diabodies, single chain (scFv) and single chain (Fv)2 (sc(Fv)2).

A "diabody" is a bivalent minibody constructed by gene fusion (see, for example, Holliger, P. et al. Proc. Natl. Acad. Sci. USA, 90:6444-6448 ( 1993), EP 404,097, WO 93/11161). Diabodies are dimers composed of two polypeptide chains. The VL and VH domains of each polypeptide chain of the diabody are joined by a linker. The number of amino acid residues that make up the linker can be 2-12 residues (eg, 3-10 residues, or 5 or about 5 residues). The linkers of the polypeptides in diabodies are typically too short to allow VL and VH to join together. Thus, VL and VH encoded by the same polypeptide chain cannot form single chain variable region fragments, but instead form dimers with different single chain variable region fragments. As a result, diabodies have two antigen-binding sites.

scFv are single-chain polypeptide antibodies obtained by linking VH and VL with a linker (see, eg, Huston et al. Proc. Natl. Acad. Sci. USA, 85:5879-5883 (1988), and Plickthun, "The Pharmacology of Monoclonal Antibodies" Vol. 113, Ed Resenburg and Moore, Springer Verlag, New York, pp. 269-315, (1994)). The order of VH and VL to be linked is not particularly limited, and can be arranged in any order. Arrangement examples include [VH]linker[VL], or [VL]linker[VH]. The H and L chain V regions in ScFv can be derived from any antibody that binds to human PD-1 or human PD-L1, or an antigen-binding fragment thereof, as described herein.

sc(Fv)2 is a minibody in which two VH and two VL are joined by a linker to form a single chain (Hudson, et al., J. Immunol. Methods, (1999) 231:177 -189 (1999)). sc(Fv)2 can be prepared, for example, by connecting scFv with a linker. The sc(Fv)2 of the present disclosure are preferably antibodies in which the 2 VHs and 2 VLs are arranged in the following order: VH, VL, VH, and VL starting from the N-terminus of the single-chain polypeptide ( [VH] linker [VL] linker [VH] linker [VL]), provided that the order of the two VHs and the two VLs is not limited to the above arrangement, they can be arranged in any order .

Bispecific Antibodies Bispecific antibodies are antibodies that have binding specificities for at least two different epitopes. Exemplary bispecific antibodies can bind to two different epitopes of the PD-1 protein. Other such antibodies may combine a PD-1 binding site with a binding site for another protein. Bispecific antibodies are prepared as full length antibodies or lower molecular weight forms thereof (e.g. F(ab') ₂ bispecific antibodies, sc(Fv)2 bispecific antibodies, diabodies bispecific antibodies) can do.

Traditional production of full-length bispecific antibodies is based on the co-expression of two immunoglobulin heavy-light chain pairs, which have different specificities (Millstein et al., Nature, 305 : 537-539 (1983)). According to a different approach, antibody variable domains with the desired binding specificities are fused to immunoglobulin constant domain sequences. DNAs encoding the immunoglobulin heavy chain fusions, and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host cell. This provides greater flexibility in adjusting the ratio of the three polypeptide fragments. However, if expression of at least two polypeptide chains in equal ratios results in high yields, it is possible to insert the coding sequences for two or all three polypeptide chains into one expression vector. .

According to another approach described in U.S. Pat. No. 5,731,168, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimer recovered from recombinant cell culture. good. A preferred interface comprises at least a portion of a _CH3 domain. In this method, one or more small amino acid side chains from the interface of the first antibody molecule are replaced with larger side chains (eg tyrosine or tryptophan). By replacing large amino acid side chains with smaller amino acid side chains (e.g., alanine or threonine), compensating "cavities" of the same or similar size as the large side chain(s) are created on the interface of the second antibody molecule. produced. This provides a mechanism for increasing the yield of the heterodimer over other unwanted end-products such as homodimers.

Bispecific antibodies include cross-linked or "heteroconjugate" antibodies. For example, one of the antibodies in the heteroconjugate can be coupled to avidin, the other to biotin. Heteroconjugate antibodies may be made using any convenient cross-linking methods.

"Diabody" technology offers an alternative mechanism for the production of bispecific antibody fragments. These fragments contain a VH connected to a VL by a linker too short to allow pairing between the two domains on the same chain. Thus, the VH and VL domains of one fragment are paired with the complementary VL and VH domains of another fragment, thereby forming two antigen-binding sites.

Multivalent Antibodies Multivalent antibodies can be internalized (and/or catabolized) faster than bivalent antibodies by cells expressing the antigen to which the antibody binds. Antibodies provided herein may be multivalent antibodies (e.g., tetravalent antibodies) having three or more antigen-binding sites, which may be due to recombination of nucleic acids encoding the polypeptide chains of the antibody. It can be easily produced by expression. A multivalent antibody can comprise dimerization domains and three or more antigen binding sites. Exemplary dimerization domains comprise (or consist of) an Fc region or a hinge region. A multivalent antibody can comprise (or consist of) from 3 to about 8 (eg, 4) antigen binding sites. A multivalent antibody optionally comprises at least one polypeptide chain (eg, at least two polypeptide chains), where the polypeptide chain(s) comprises two or more variable domains. For example, the polypeptide chain(s) may comprise VD1-(X1) _n -VD2-(X2) _n -Fc, where VD1 is the first variable domain and VD2 is the second variable domain where Fc is one polypeptide chain of the Fc region, X1 and X2 represent amino acids or peptide spacers, and n is 0 or 1.

Conjugated Antibodies The antibodies disclosed herein may be conjugated to polymers such as polyethylene glycol (PEG), polyethyleneimine (PEI) modified with PEG (PEI-PEG), polyglutamic acid (PGA) (N-(2- hydroxypropyl) methacrylamide (HPMA) copolymers), hyaluronic acid, radioactive substances (e.g. ⁹⁰ Y, ¹³¹ I) fluorescent substances, luminescent substances, haptens, enzymes, metal chelates, drugs and toxins (e.g. Conjugated antibodies may be conjugated to a variety of molecules, including macromolecular substances such as exotoxin A, ricin (eg, deglycosylated ricin A chain)).

In one embodiment, to improve the cytotoxicity of antibodies that bind human PD-1 or human PD-L1, and thus improve their therapeutic efficacy, the antibodies are treated with a radioisotope and a cytotoxic agent. conjugated with highly toxic substances, including These conjugates can selectively deliver a toxic load to target sites (ie, cells expressing antigens recognized by antibodies), while cells not recognized by antibodies are spared. To minimize toxicity, conjugates are generally engineered based on molecules with short serum half-lives (hence the use of mouse sequences and IgG3 or IgG4 isotypes).

In certain embodiments, the antibody, or antigen-binding fragment thereof, that binds to human PD-1 or human PD-L1 is, eg, in blood, serum, or other tissue, eg, at least 1.5, 2, 5 , 10, or 50-fold, with moieties that improve its stabilization and/or retention in circulation. For example, an antibody, or antigen-binding fragment thereof, that binds human PD-1 or human PD-L1 is associated (e.g., conjugated) with a polymer, e.g., a substantially non-antigenic polymer such as a polyalkylene oxide or polyethylene oxide can do. Suitable polymers vary substantially by weight. Polymers having molecular number average weights in the range of about 200 to about 35,000 Daltons (or about 1,000 to about 15,000, and 2,000 to about 12,500) can be used. For example, an antibody, or antigen-binding fragment thereof, that binds human PD-1 or human PD-L1 can be conjugated to a water-soluble polymer, eg, a hydrophilic polyvinyl polymer such as polyvinyl alcohol or polyvinylpyrrolidone. Examples of such polymers include polyalkylene oxide homopolymers such as polyethylene glycol (PEG) or polypropylene glycol, polyoxyethylenated polyols, copolymers thereof and block copolymers thereof, provided that the water solubility of the block copolymers is maintained. provided that Additional useful polymers include polyoxyalkylenes such as polyoxyethylene, polyoxypropylene, and block copolymers of polyoxyethylene and polyoxypropylene, polymethacrylates, carbomers, and branched or unbranched polysaccharides. .

Conjugate antibodies as described above can be prepared by chemically modifying the antibodies described herein or low molecular weight forms thereof. Methods for modifying antibodies are well known in the art (eg US5057313 and US5156840).

Methods of Producing Antibodies Antibodies can be produced in bacterial or eukaryotic cells. Some antibodies, such as Fab', can be used in bacterial cells such as E. coli. can be produced in E. coli cells. Antibodies can also be produced in eukaryotic cells, such as transformed cell lines (eg CHO, 293E, COS). Additionally, antibodies (eg, scFv') can be expressed in yeast cells such as Pichia (eg, Powers et al., J Immunol Methods. 251:123-35 (2001)), Hanseula, or Saccharomyces. To produce the antibody of interest, a polynucleotide encoding the antibody is constructed, introduced into an expression vector, and expressed in a suitable host cell. Standard molecular biology techniques are used to prepare recombinant expression vectors, transfect host cells, select transformants, culture host cells, and recover antibodies.

If the antibody is to be expressed in bacterial cells (eg, E. coli), the expression vector should have characteristics that allow amplification of the vector within the bacterial cells. In addition, E. coli, such as JM109, DH5α, HB101, or XL1-Blue. When E. coli is used as a host, the vector contains promoters such as the lacZ promoter (Ward et al., 341:544-546 (1989), the araB promoter (Better et al., Science, 240:1041-1043 (1988)). ), or must have a T7 promoter capable of enabling efficient expression in E. coli Examples of such vectors include, for example, M13 series vectors, pUC series vectors, pBR322, pBluescript, pCR-Script, pGEX-5X-1 (Pharmacia), "QIAexpress system" (QIAGEN), pEGFP, and pET (when this expression vector is used, the host is preferably BL21 expressing T7 RNA polymerase). The expression vector may contain a signal sequence for antibody secretion: for production into the periplasm of E. coli, the pelB signal sequence (Lei et al., J. Bacteriol., 169:4379 ( 1987)) may be used as signal sequences for antibody secretion For bacterial expression, the calcium chloride method or the electroporation method may be used to introduce the expression vector into bacterial cells.

When the antibody is expressed in animal cells, such as CHO, COS, and NIH3T3 cells, the expression vector should contain promoters necessary for expression in these cells, such as the SV40 promoter (Mulligan et al. Nature, 277:108). (1979)), MMLV-LTR promoter, EF1α promoter (Mizushima et al., Nucleic Acids Res., 18:5322 (1990)), or CMV promoter. In addition to the nucleic acid sequence encoding an immunoglobulin or domain thereof, the recombinant expression vector carries additional sequences, such as sequences that regulate the replication of the vector in a host cell (e.g., origin of replication) and selectable marker genes. can be made The selectable marker gene can facilitate selection of host cells into which the vector has been introduced (e.g., US Pat. Nos. 4,399,216, 4,634,665, and 5,179,017). (see ). For example, typically the selectable marker gene confers resistance to drugs, such as G418, hygromycin, methotrexate, on a host cell into which the vector has been introduced. Examples of vectors with selectable markers include pMAM, pDR2, pBK-RSV, pBK-CMV, pOPRSV, and pOP13.

In one embodiment, antibodies are produced in mammalian cells. Exemplary mammalian host cells for expressing antibodies include Chinese Hamster Ovary (CHO cells) (dhfr-, as described in Urlaub and Chasin (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220 ⁾ . human embryonic kidney 293 cells (e.g., 293, 293E), including CHO cells and used with the DHFR selectable marker, e.g., as described in Kaufman and Sharp (1982) Mol. , 293T), COS cells, NIH3T3 cells, lymphocytic cell lines such as NSO myeloma cells and SP2 cells, and transgenic animals, such as cells derived from transgenic mammals. For example, the cells are mammary epithelial cells.

In an exemplary system for antibody expression, a recombinant expression vector encoding both the antibody heavy and light chains of an antibody that binds a human PD-1 or human PD-L1 antibody (e.g., Antibody X) is It is introduced into dhfr ⁻ CHO cells by calcium phosphate-mediated transfection. Within the recombinant expression vector, the antibody heavy and light chain genes are each linked to an enhancer/promoter regulatory element (e.g., SV40, such as the CMV enhancer/AdMLP promoter regulatory element or the SV40 enhancer/AdMLP promoter regulatory element, CMV, adenovirus, etc.). origin) and drives high-level transcription of the gene. The recombinant expression vector also carries a DHFR gene that allows selection of CHO cells transfected with the vector using methotrexate selection/amplification. The selected transformed host cells are cultured to permit expression of the antibody heavy and light chains, and the antibody is recovered from the culture medium.

Antibodies can also be produced by transgenic animals. For example, US Pat. No. 5,849,992 describes methods of expressing antibodies in the mammary gland of transgenic mammals. A transgene is constructed containing a milk-specific promoter and nucleic acid encoding the antibody of interest and a signal sequence for secretion. The milk produced by females of such transgenic mammals contains the antibody of interest secreted therein. Antibodies can be purified from milk or used directly for some applications. Animals containing one or more of the nucleic acids described herein are also provided.

Antibodies of the present disclosure can be isolated from the inside or outside (such as the medium) of host cells and purified as substantially pure and homogeneous antibodies. Isolation and purification methods commonly used for antibody purification may be used for antibody isolation and purification, and are not limited to any particular method. Antibodies are suitably subjected to, for example, column chromatography, filtration, ultrafiltration, salting out, solvent precipitation, solvent extraction, distillation, immunoprecipitation, SDS-polyacrylamide gel electrophoresis, electrofocusing, dialysis, and recrystallization. It can be isolated and purified by selection and combination. Chromatography includes, for example, affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration, reverse phase chromatography, and adsorption chromatography (Strategies for Protein Purification and Characterization: A Laboratory Course Manual. Ed Daniel R.; Marshak et al., Cold Spring Harbor Laboratory Press, 1996). Chromatography can be performed using liquid phase chromatography such as HPLC and FPLC. Columns used for affinity chromatography include protein A columns and protein G columns. Examples of columns using protein A columns include Hyper D, POROS, and Sepharose FF (GE Healthcare Biosciences). The disclosure also includes antibodies that are highly purified using these purification methods.

Antibodies with Altered Glycosylation Different glycoforms can profoundly affect the properties of therapeutic agents, including pharmacokinetics, pharmacodynamics, receptor interactions, and tissue-specific targeting (Graddis et al., 2002, Curr. Pharm Biotechnol. 3:285-297). In particular, for antibodies, oligosaccharide structures, in addition to their effector functions (e.g., binding to complement complex C1, which induces CDC, and binding to FcγR receptors, which are responsible for regulating the ADCC pathway), are protease resistant. , may affect the serum half-life of antibodies mediated by FcRn receptors, phagocytosis, and properties related to antibody feedback (Nose and Wigzell, 1983; Leatherbarrow and Dwek, 1983; Leatherbarrow et al., 1985; Walker et al., 1989, Carter et al., 1992, PNAS, 89:4285-4289).

Thus, another means of modulating antibody effector functions involves altering the glycosylation of the antibody constant region. Altered glycosylation includes, for example, decreasing or increasing the number of glycosylated residues, changing the pattern or position of glycosylated residues, and altering sugar structure(s). The oligosaccharides found in human IgG influence the extent of their effector function (Raju, T.S. BioProcess International April 2003.44-53), and the microheterogeneity of human IgG oligosaccharides contributes to CDC and ADCC. binding to various Fc receptors, as well as binding to Clq proteins (Wright A. & Morrison SL. TIBTECH 1997, 15 26-32, Shields et al. J Biol Chem 2001 276(9):6591-604, Shields et al.J Biol Chem.2002, 277(30):26733-40, Shinkawa et al.J Biol Chem.2003 278(5):3466-73, Umana et al. al., Nat Biotechnol.1999 Feb, 17(2):176-80). For example, the ability of IgG to bind C1q and activate the complement cascade depends on the presence, absence, or modification of a carbohydrate moiety located between the two CH2 domains (usually anchored at Asn297). (Ward and Ghetie, Therapeutic Immunology 2:77-94 (1995).

Glycosylation sites within Fc-containing polypeptides, eg, antibodies, such as IgG antibodies, can be identified by standard techniques. Identification of glycosylation sites may be based on experimental or sequence analysis or modeling data. Consensus motifs, ie, amino acid sequences recognized by various glycosyltransferases, have been described. For example, the consensus motif for N-linked glycosylation motifs is frequently NXT or NXS, where X can be any amino acid except proline. Several algorithms for finding potential glycosylation motifs have also been described. Accordingly, to identify potential glycosylation sites within an antibody or Fc-containing fragment, the sequence of the antibody is analyzed using, for example, public databases such as the website provided by the Center for Biological Sequence Analysis. Examine (see NetNGlyc service for predicting N-linked glycosylation sites and NetOGlyc service for predicting O-linked glycosylation sites).

In vivo studies confirm the reduced effector function of aglycosyl antibodies. For example, aglycosyl anti-CD8 antibodies fail to deplete CD8-bearing cells in mice (Isaacs, 1992 J. Immunol. 148:3062), and aglycosyl anti-CD3 antibodies induce cytokine release syndrome in mice or humans. (Boyd, 1995 (supra), Friend, 1999 Transplantation 68:1632). Aglycosylated forms of PD-1 antibodies also have reduced effector functions.

Importantly, while removal of glycans within the CH2 domain appears to significantly affect effector function, other functional and physical properties of the antibody remain unchanged. Specifically, removal of glycans has been shown to have little or no effect on serum half-life and binding to antigen (Nose, 1983 (supra); Tao, 1989 (supra); Dorai, 1991 (supra), Hand, 1992 (supra), Hobbs, 1992 (supra), Mol. Immunol. 29:949).

Antibodies that bind human PD-1 or human PD-L1 of the present disclosure are modified or can be modified. Methods for modifying the glycosylation sites of antibodies are described, for example, in US Pat. No. 6,350,861 and US Pat. Alternatively, antibodies of the present disclosure can be produced with reduced or no glycosylation.

Solid Tumors and Cancer The methods described herein involve the treatment of cancer, preferably solid tumors.

In some embodiments, the solid tumor is skin cancer, lung cancer, lymphoma, sarcoma, bladder cancer, ureteral cancer, urethral cancer, and urachal cancer, gastric cancer, cervical cancer, liver cancer, breast cancer, renal cancer, Selected from squamous cell carcinoma, colon cancer, endometrial cancer, anal cancer, and tumors with high microsatellite instability (MSI-H), mismatch repair deficiency (dMMR), and DNA polymerase epsilon exonuclease domain mutation-positive disease be done.

In some embodiments, the solid tumor is cholangiocarcinoma, melanoma, non-small cell lung cancer, small cell lung cancer, Hodgkin's lymphoma, urothelial carcinoma, gastric cancer, hepatocellular carcinoma, Merkel cell carcinoma, triple negative breast cancer, renal selected from cell carcinoma, squamous cell carcinoma of the head and neck, and colon cancer.

In some embodiments, the solid tumor is microsatellite stable (MSS). In some embodiments, the solid tumor is PD-L1 positive. In some embodiments, the solid tumor is microsatellite stable (MSS) and PD-L1 positive. In some embodiments, the solid tumor is endometrial cancer (eg, endometrial carcinoma). In some embodiments, the solid tumor is bladder cancer (eg, non-muscle invasive bladder cancer, such as Bacillus Calmette-Guerin non-muscle invasive bladder cancer).

Examples of cancers treatable using the treatment methods and regimens of the present disclosure include, but are not limited to, bone cancer, spleen cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer. , ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, carcinoma of the endometrium, endometrial cancer, cervical cancer , vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, ureteral cancer, urethral cancer, urachal cancer, chronic or acute leukemia including gastric cancer, penile cancer, acute myeloid leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney Cancer or urethral carcinoma, renal pelvic carcinoma, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell Included are cancers, T-cell lymphomas, environmentally induced cancers including asbestos-induced cancers, and combinations of the above cancers. The disclosed methods are also useful for treating metastatic cancers, particularly metastatic cancers that express PD-L1.

In some embodiments, the cancer is endometrial cancer. In some embodiments, the endometrial cancer is microsatellite stable (MSS). In some embodiments, the endometrial cancer is PD-L1 positive. In some embodiments, the endometrial cancer is microsatellite stable (MSS) and PD-L1 positive. In some embodiments, the endometrial cancer is metastatic endometrial cancer. In some embodiments, the endometrial cancer is metastatic, microsatellite-stable (MSS), and PD-L1-positive endometrial cancer (e.g., metastatic, microsatellite-stable (MSS), and PD-L1-positive endometrial cancer). L1-positive endometrial cancer).

In some embodiments, the present application treats a patient with microsatellite stable (MSS), PD-L1 positive endometrial cancer (e.g., microsatellite stable (MSS), PD-L1 positive endometrial cancer) A method, comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 600 mg on a free base basis twice daily;
(ii) an antibody, or antigen-binding fragment thereof, that binds to human PD-1, wherein the antibody comprises (ii-1) variable heavy (VH) complementarity determining regions (CDR) 1, VH CDR2, and VH CDR3; administering an antibody, or antigen-binding fragment thereof, comprising a VH domain and (ii-2) a VL domain comprising variable light (VL) CDR1, VL CDR2, and VL CDR3;
(a) the VH CDR1 comprises the amino acid sequence SYWMN (SEQ ID NO: 6);
(b) VH CDR2 comprises the amino acid sequence VIHPSDSETWLDQKFKD (SEQ ID NO: 7);
(c) the VH CDR3 comprises the amino acid sequence EHYGTSPFAY (SEQ ID NO: 8);
(d) the VL CDR1 comprises the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 9);
(e) VL CDR2 comprises the amino acid sequence AASNQGS (SEQ ID NO: 10);
(f) the VL CDR3 comprises the amino acid sequence QQSKEVPYT (SEQ ID NO: 11);
Methods are provided herein wherein the antibody is administered at a fixed dose of about 375 mg once every three weeks, or about 500 mg once every four weeks. In some embodiments, the microsatellite stable (MSS), PD-L1 positive endometrial cancer is metastatic microsatellite stable (MSS), PD-L1 positive endometrial cancer.

In some embodiments, the cancer is bladder cancer. In some embodiments, the bladder cancer is non-muscle invasive bladder cancer (eg, Bacillus Calmette-Guerin non-responsive non-muscle invasive bladder cancer).

In some embodiments, the application provides a method of treating non-muscle invasive bladder cancer in a patient, comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 600 mg on a free base basis twice daily;
(ii) an antibody, or antigen-binding fragment thereof, that binds to human PD-1, wherein the antibody comprises (ii-1) variable heavy (VH) complementarity determining regions (CDR) 1, VH CDR2, and VH CDR3; administering an antibody, or antigen-binding fragment thereof, comprising a VH domain and (ii-2) a VL domain comprising variable light (VL) CDR1, VL CDR2, and VL CDR3;
(a) the VH CDR1 comprises the amino acid sequence SYWMN (SEQ ID NO: 6);
(b) VH CDR2 comprises the amino acid sequence VIHPSDSETWLDQKFKD (SEQ ID NO: 7);
(c) the VH CDR3 comprises the amino acid sequence EHYGTSPFAY (SEQ ID NO: 8);
(d) the VL CDR1 comprises the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 9);
(e) VL CDR2 comprises the amino acid sequence AASNQGS (SEQ ID NO: 10);
(f) the VL CDR3 comprises the amino acid sequence QQSKEVPYT (SEQ ID NO: 11);
Methods are provided herein wherein the antibody is administered at a fixed dose of about 375 mg once every three weeks, or about 500 mg once every four weeks.

In some embodiments, the bladder cancer is Bacillus Calmette-Guerin non-responsive non-muscle invasive bladder cancer (ie, BCG non-responsive non-muscle invasive bladder cancer). In some embodiments, the bladder cancer is high-risk BCG non-responsive non-muscle invasive bladder cancer. In some embodiments, the bladder cancer is high-risk BCG non-responsive non-muscle invasive bladder cancer (eg, with or without papillary tumors) with carcinoma in situ (CIS). In some embodiments, patients with non-muscle invasive bladder cancer are ineligible or selected not to undergo cystectomy.

In some embodiments, cancers treatable by the methods of the present disclosure have microsatellite instability-high (MSI-H), mismatch repair deficient (dMMR), or DNA polymerase epsilon exonuclease domain mutation-positive disease. Includes tumors.

In some embodiments, the cancer has a ratio of indoleamine-2,3-dioxygenase (IDO) to tryptophan-2,3-dioxygenase (TDO) of at least 10.

In some embodiments, the cancer has an indoleamine-2,3-dioxygenase-high (IDOhi) to tryptophan-2,3-dioxygenase-low (TDOlow) ratio of at least 50%.

In some embodiments, the cancer is cervical cancer.

In some embodiments, the cancer is kidney cancer.

In some embodiments, the cancer is renal clear cell carcinoma.

In some embodiments, the cancer is lung cancer.

In some embodiments, the cancer is adenocarcinoma of the lung.

In some embodiments, the cancer is squamous cell carcinoma of the lung.

In some embodiments, the cancer is non-small cell lung cancer.

In some embodiments, the cancer is head and neck cancer.

In some embodiments, the cancer is head and neck squamous cell carcinoma.

In some embodiments, cancers treatable by the methods of the present disclosure include melanoma (e.g., metastatic malignant melanoma), renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone refractory cancer). prostate adenocarcinoma), breast cancer, colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), head and neck squamous cell carcinoma, urothelial carcinoma (e.g., bladder), and cancers with high microsatellite instability (MSIhigh) is included. Further, the present disclosure includes refractory or recurrent malignancies whose growth may be inhibited using the methods of the present disclosure.

In some embodiments, cancers treatable using the methods of the present disclosure include solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, renal cancer). , liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), blood cancers (e.g., lymphoma, acute lymphoblastic leukemia (ALL), acute bone marrow chronic lymphocytic leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and recurrent follicular), Hodgkin lymphoma or multiple lymphomas such as myeloma, leukemia) as well as combinations of said cancers.

In some embodiments, cancers treatable using the treatment methods and regimens of the present disclosure include, but are not limited to, cholangiocarcinoma, bil duct cancer, biliary tract cancer, triple Negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing sarcoma, brain cancer, brain tumor, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, pancreatic islet cell cancer, oral cancer, mouth cancer, throat cancer , laryngeal cancer, lip cancer, mesothelioma, neck cancer, nasal cavity cancer, ocular cancer, ocular melanoma, pelvic cancer, rectal cancer, renal cell cancer, salivary gland cancer, sinus cancer, spinal cord cancer, tongue Included are carcinoma, tubular carcinoma, urethral carcinoma, and ureteral carcinoma.

In some embodiments, diseases and indications treatable using the treatment methods and regimens of the present disclosure include blood cancer, sarcoma, lung cancer, gastrointestinal cancer, urogenital cancer, liver cancer, bone cancer, neurological Including, but not limited to, systemic cancers, gynecologic cancers, and skin cancers.

Exemplary blood cancers include lymphomas and leukemias such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL). , chronic myelogenous leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and recurrent follicular), Hodgkin's lymphoma, myeloproliferative disease (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), post-essential thrombocytemia myelofibrosis transitioning from essential thrombocythemia, myelofibrosis transitioning from polycythemia vera ( post polycythemia vera-myelofibrosis), myelofibrosis transitioned from polycythemia vera/essential thrombocythemia, and essential thrombocythemia (ET)), myelodysplastic syndrome (MDS), T-cell acute lymphoblastic Examples include lymphoma (T-ALL) and multiple myeloma (MM).

Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, Askin's tumor, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, Lipoma, harmatoma, teratoma, grape sarcoma, chondrosarcoma, malignant hemangioendothelioma, malignant schwannoma, alveolar soft part sarcoma, phyllodes cyscoma, dermatofibrosarcoma protuberans, tendonoid, desmoplastic small round Cellular tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, lymphangiosarcoma, lymphosarcoma, malignant They include peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, synovial sarcoma, and undifferentiated pleomorphic sarcoma.

Exemplary lung cancers include non-small cell lung cancer (NSCLC) (e.g., squamous cell NSCLC), small cell lung cancer, bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), Alveolar (bronchial) carcinoma, bronchial adenoma, cartilaginous hamartoma, and mesothelioma.

Exemplary gastrointestinal cancers include esophageal cancer (carcinoma, squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma, adenocarcinoma), pancreatic cancer (ductal adenocarcinoma, islet cell tumor, glucagon-producing tumor, gastrin-producing tumor, carcinoid tumor, VIP-producing tumor), small intestinal cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), Included are large bowel cancers (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) and colorectal cancer (colonic adenocarcinoma).

Exemplary urogenital tract cancers include kidney cancer (adenocarcinoma, Wilms tumor, [nephroblastoma]), bladder and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma). carcinoma, sarcoma), and testicular cancer (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma). In some embodiments, the cancer is a cancer of the urinary system (eg, papillary renal carcinoma, testicular germ cell carcinoma, chromophobe renal cell carcinoma, clear cell renal carcinoma, or prostate adenocarcinoma).

Exemplary liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular sarcoma), multiple myeloma, malignant melanoma. Cellular tumors include chordoma, osteochondroma (osteochondral exostosis), benign cartilage tumor, chondroblastoma, chondromyxofibroma, osteoid, and giant cell tumors.

Exemplary nervous system cancers include skull (osteoma, hemangioma, granuloma, xanthoma, osteitis osteoarthritis), meninges (meningioma, meningosarcoma, glioma), brain (astatellous). Cell tumor, medulloblastoma, glioma, ependymoma, germinoma (pineocytoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital cancer), and cancer of the spinal cord (neurofibroma, meningioma, glioma, sarcoma), as well as neuroblastoma and Lhermitte-Dacros disease.

Exemplary gynecologic cancers include uterine (endometrial cancer), cervical (cervical cancer, preneoplastic cervical dysplasia), ovarian (ovarian cancer (serous cystadenocarcinoma, serous adenocarcinoma, mucinous cystadenocarcinoma, unclassifiable carcinoma), granulosa-capsular cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosis) sarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, sarcoma grape (embryonal rhabdomyosarcoma), and fallopian tube (carcinoma).

Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma (e.g. cutaneous squamous cell carcinoma), Kaposi's sarcoma, mole dysplasia nevus, lipoma, hemangioma, dermatofibroma, and keloid. is mentioned. In some embodiments, diseases and indications treatable using the treatment methods and regimens of the present disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, cholangiocarcinoma, esophageal cancer, and urothelial cancer.

In some embodiments, diseases and indications treatable using the treatment methods and regimens of the present disclosure include, but are not limited to, adrenal gland tumors, AIDS-related cancers, alveolar soft tissue sarcoma, astrocytic tumors, bladder cancer. , bone cancer, brain and spinal cord cancer, metastatic brain tumor, breast cancer, carotid body tumor, cervical cancer, chondrosarcoma, chordoma, pigmented renal cell carcinoma, clear cell carcinoma, colon cancer, colorectal cancer, benign fibrous skin histocytoma, degenerative small round cell tumor, epithelioma, Ewing tumor, extraosseous myxoid chondrosarcoma, desmoplastic bone, dysplasia of bone fibrosis, gallbladder or cholangiocarcinoma, gastric cancer, gestational trophoblastic disease, Germ cell tumor, head and neck cancer, hepatocellular carcinoma, islet cell tumor, Kaposi's sarcoma, renal cancer, leukemia, lipoma/benign lipoma, liposarcoma/malignant lipoma, liver cancer, lymphoma, lung cancer, medulloblastoma, melanoma , meningioma, multiple endocrine tumors, multiple myeloma, myelodysplastic syndrome, neuroblastoma, neuroendocrine tumor, ovarian cancer, pancreatic cancer, papillary thyroid cancer, parathyroid cancer, childhood cancer, peripheral nerve sheath Tumor, chromogenic cell tumor, pituitary tumor, prostate cancer, secondary uveal melanoma, rare hematologic disease, renal metastatic carcinoma, rhabdomoma, rhabdomyosarcoma, sarcoma, skin cancer, soft tissue sarcoma, squamous cell cancer, gastric cancer, synovial sarcoma, testicular cancer, thymic carcinoma, thymoma, metastatic thyroid cancer, and uterine cancer.

In some embodiments, the cancer treatment methods and regimens of the present disclosure are used for colon cancer, hepatocellular carcinoma, glioma, renal cancer, breast cancer, multiple myeloma, bladder cancer, neuroblastoma, sarcoma, Non-Hodgkin's lymphoma, non-small cell lung cancer, ovarian cancer, pancreatic cancer, rectal cancer, acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute B-lymphoblastic leukemia (B-ALL), chronic lymphocytic Non-Hodgkin's lymphomas (NHL), including leukemia (CLL), hairy cell leukemia (HCL), spore-forming plasma cell dendritic cell tumor (BPDCN), mantle cell leukemia (MCL) and small lymphocytic lymphoma (SLL), Hodgkin It is selected from, but not limited to, lymphoma, systemic mastocytosis, and Burkitt's lymphoma.

As used herein, the term "cell" means a cell that is in vitro, ex vivo or in vivo. In some embodiments, an ex vivo cell can be part of a tissue sample excised from an organism such as a mammal. In some embodiments, an in vitro cell can be a cell in cell culture. In some embodiments, an in vivo cell is a cell that lives in an organism such as a mammal.

As used herein, the term "contacting" refers to bringing the indicated moieties together in an in vitro or in vivo system. For example, "contacting" an IDO enzyme with epacadostat includes administering epacadostat to an individual, such as a human, or a patient having IDO, as well as, for example, a cell preparation or purified preparation containing the IDO enzyme. Including introducing epacadostat to the containing sample.

As used herein, the terms "subject," "individual," or "patient" are used interchangeably and refer to mammals, preferably mice, rats, other rodents, rabbits, dogs, It refers to any animal, including cats, pigs, cows, sheep, horses, or primates, and most preferably humans.

As used herein, the term "treating" or "treatment" refers to: 1) inhibiting a disease, e.g. , inhibit the condition or disorder (i.e., prevent further development of the pathology and/or symptomology), or 2) ameliorate the disease, e.g., experience or exhibit the pathology or symptomology of the disease, condition or disorder. Refers to ameliorating a disease, condition or disorder (ie, reversing the medical condition and/or symptomatology) in an individual suffering from disease.

As used herein, the term "prevent" or "prophylaxis" refers to individuals who may be predisposed to a disease, condition or disorder but who have not yet experienced or exhibited disease pathology or symptomology. refers to preventing a disease, condition or disorder of

Anal Canal Squamous Cell Carcinoma Anal canal squamous cell carcinoma (SCAC) accounts for nearly 3% of gastrointestinal cancers and is increasing in frequency due to its association with HPV and HIV infection. Although most patients have localized disease, about 25% of patients develop systemic metastases, and these individuals have a poor 5-year survival rate. Salvage chemotherapy with platinum-based regimens is an accepted standard of care, but responses have not been sustained, and progression-free and overall survival following these treatments is measured only in months. There are no acceptable salvage treatments for patients who progress after first-line chemotherapy.

Merkel Cell Carcinoma Merkel cell carcinoma is a rare, aggressive cutaneous malignancy due to multiple factors, including Merkel cell polyomavirus, UV irradiation, and immunosuppression. The disease is typically seen in the elderly with fair skin types and has a poor prognosis and poor survival compared to other cutaneous malignancies. Surgery and/or radiation therapy are indicated, and potential treatments for local disease and recurrence are common.

The 5-year survival rates for MCC patients are 75%, 59%, and 25% for primary localized tumors, tumors with regional lymph node metastasis (or local recurrence), and tumors with distant metastasis, respectively. More than 30% of patients develop distant metastatic disease, and these patients have a 5-year survival rate of only about 10%.

Historically, metastatic MCC has been treated with chemotherapy regimens similar to those used for small cell lung cancer. Platinum-based chemotherapy provides high initial response rates of short duration. A survival advantage of chemotherapy in this disease has not been demonstrated. Chemotherapy is also associated with severe toxicity and risk of toxic death, especially in elderly patients.

Endometrial Cancer Endometrial cancer is the fourth most common cancer affecting American women, with an estimated 60,050 new cases diagnosed and an estimated 10,470 endometrial cancer-related cases deaths have occurred, making it the sixth most common cancer-related death affecting American women. Worldwide, it is the fourth most common cause of cancer-related death in women. Endometrial cancer is the most common gynecologic malignancy afflicting women, and adenocarcinoma is the most common histology. Early-diagnosed cancers provide a good prognosis with curative options of surgery and/or radiation, whereas aggressive late-stage cancers have limited curative treatment options and a 5-year survival rate of 20. ~60%. Standard treatments for locally advanced or metastatic cancer include systemic treatments such as hormonal therapy, single agent chemotherapy such as doxorubicin, or platinum-based combination chemotherapy regimens such as carboplatin and docetaxel. Due to the poor long-term prognosis of these patients, additional and new treatments are required.

Pharmaceutical Compositions In some embodiments, the compound epacadostat can be formulated as part of a pharmaceutical composition. In some embodiments, antibodies that bind human PD-1 or human PD-L1 can be formulated as part of a pharmaceutical composition. A pharmaceutical composition comprising a compound described herein and an antibody, or antigen-binding fragment thereof, that binds to human PD-1 or human PD-L1 treats, for example, a disorder described herein To that end, it can be formulated as a pharmaceutical composition for administration to a subject. Typically, a pharmaceutical composition includes a pharmaceutically acceptable carrier. As used herein, a "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents that are physiologically compatible. including drugs. The composition can include pharmaceutically acceptable salts, such as acid or base addition salts (see, eg, Berge, SM, et al. (1977) J. Pharm. Sci. 66: 1-19).

Pharmaceutical formulation is a well-established art, see, for example, Gennaro (ed.), Remington: The Science and Practice of Pharmacy, 20th ^ed . , Lippincott, Williams & Wilkins (2000) (ISBN: 0683306472), Ansel et al. , Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ^Ed . , Lippincott Williams & Wilkins Publishers (1999) (ISBN: ^0683305727 ), and Kibbe (ed.), Handbook of Pharmaceutical Excipients American Pharmaceutical Association, 3. (2000) (ISBN: 091733096X).

Pharmaceutical compositions can be in various forms. These include liquid, semi-solid and solid dosage forms such as, for example, liquid solutions (eg, injection and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. . The preferred form will depend on the intended mode of administration and therapeutic use. Typically, compositions for the agents described herein are in the form of injectable or infusible solutions.

The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for stable storage at high concentrations. Sterile injectable solutions incorporate one or a combination of the above-listed ingredients in the required amount of an agent described herein in an appropriate solvent, optionally followed by filter sterilization. can be prepared by Generally, dispersions are prepared by incorporating the agent described herein into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. . In the case of sterile powders for the preparation of sterile injectable solutions, the preferred method of preparation is to combine the agents described herein with any additional desired ingredients from previously sterile-filtered solutions. Vacuum-drying and freeze-drying, which result in a powder loaded with the desired ingredients. Proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prolonged absorption of an injectable composition can be brought about by including in the composition an agent that delays absorption, such as monostearate and gelatin.

In certain embodiments, an antibody, or antigen-binding fragment thereof, that binds human PD-1 or human PD-L1 is combined with a carrier that protects the compound against rapid release, such as sustained release formulations and microencapsulated delivery systems, including implants. can be prepared using Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known. See, for example, Sustained and Controlled Release Drug Delivery Systems, J. Am. R. Robinson, ed. Marcel Dekker, Inc.; , New York (1978).

In some embodiments, the compound is formulated as part of a pharmaceutical composition, further comprising at least one excipient.

In some embodiments, the compound is mixed with an excipient, diluted by an excipient, or packaged, for example, in a capsule, sachet, paper or other form, in making the compositions described herein. is enclosed in a carrier in the form of a container. When the excipient serves as a diluent, it can be a solid, semisolid, or liquid material that acts as a vehicle, carrier, or medium for the active ingredient. Compositions thus include tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid media), e.g., up to 10% by weight of active compound. ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

In some embodiments, the pharmaceutical compositions described herein are in tablet form.

In preparing a formulation, the compound can be milled to provide the appropriate particle size prior to combining with other ingredients. In some embodiments, the compound can be ground to a particle size of less than 200 mesh. In some embodiments, the particle size can be adjusted by milling, eg, about 40 mesh, to provide a substantially uniform distribution in the formulation.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose. , water, syrup, and methylcellulose. The formulations can additionally include lubricating agents such as talc, magnesium stearate and mineral oil, wetting agents, emulsifying and suspending agents, preserving agents such as methylbenzoate and propylhydroxybenzoate, sweetening agents and flavoring agents. The compositions described herein can be formulated to provide rapid, sustained, or delayed release of the active ingredient after administration to the patient by using procedures known in the art.

Compositions may be formulated in unit dosage form. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, each unit in association with suitable pharmaceutical excipients. , containing a predetermined amount of compound calculated to produce a desired therapeutic effect (eg, a desired PK profile).

In certain embodiments, for preparing solid compositions such as tablets, the compound is mixed with pharmaceutical excipients to form a solid preformulation composition comprising a homogeneous mixture of the compound. When these preformulation compositions are referred to as homogenous, the compound is typically dispersed evenly throughout the composition so that the composition can be divided into equally effective units such as tablets, pills and capsules. It can be easily subdivided into dosage forms. This solid preformulation is then subdivided into unit dosage forms.

The tablets or pills of the present disclosure may be coated or otherwise compounded to provide a dosage form affording the advantage of long action. For example, a tablet or pill may contain an inner dose and an outer dose component, the latter in the form of an envelope over the former. The two components can be separated by an enteric layer that resists disintegration in the stomach and allows the internal components to pass intact into the duodenum or to be delayed in release. A variety of materials are available for such enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

Liquid forms in which the compositions described herein may be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and cottonseed, sesame, coconut, or Emulsions flavored with edible oils such as peanut oil, as well as elixirs and similar pharmaceutical vehicles are included.

In some embodiments, compositions described herein can be sterilized by conventional sterilization techniques, or may be sterile filtered. Aqueous solutions can be packaged for use as is, or can be lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparation will typically be 3-11, more preferably 5-9, most preferably 7-8. It will be appreciated that the use of certain of the aforementioned excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

Combination therapy I. Cancer Therapy Cancer cell proliferation and survival can be affected by dysfunction of multiple signaling pathways. Therefore, it is useful to treat such conditions by combining different enzyme/protein/receptor inhibitors that exhibit different selectivities in targeting so as to modulate the activity of different enzyme/protein/receptor inhibitors. is. Targeting more than one signaling pathway (or two or more biological molecules involved in a given signaling pathway) reduces the likelihood of drug resistance occurring in a cell population and/or can reduce the toxicity of

In addition to one or more additional pharmaceutical agents such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressive agents, cancer immunotherapeutic agents, metabolic enzyme inhibitors, chemokine receptor inhibitors, and phosphatase inhibitors. targeted therapies such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF, FAK, CDK2, and CDK4/6 kinase inhibitors Such as, for example, those described in WO2006/056399, can be used in combination with the therapeutic methods and regimens of the present disclosure to treat cancer and or solid tumors. Other agents, such as therapeutic antibodies, can be used in combination with the therapeutic methods and regimens of this disclosure for the treatment of cancer and solid tumors. The one or more additional pharmaceutical agents can be administered to the patient concurrently or sequentially.

Treatment methods as disclosed herein include one or more other enzyme/protein/receptor inhibitors to treat diseases such as cancer and other diseases or disorders described herein. Can be used in combination with drug therapy. For example, the therapeutic methods and regimens of the present disclosure can be combined with inhibitors of one or more of the following kinases for the treatment of cancer: Akt1, Akt2, Akt3, BCL2, CDK2, CDK4/6, TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFαR, PDGFβR , PI3K (alpha, beta, gamma, delta, and multiple or alternatively), CSF1R, KIT, FLK-II, KDR/FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c- Met, PARP, Ron, Sea, TRKA, TRKB, TRKC, TAM kinases (Axl, Mer, Tyro3), FLT3, VEGFR/Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. Non-limiting examples of inhibitors that can be combined with the therapeutic methods and regimens of the present disclosure for the treatment of cancer include FGFR inhibitors (FGFR1, FGFR2, FGFR3, or FGFR4, e.g. pemigatinib (INCY54828), INCB62079 ), EGFR inhibitors (also known as ErB-1 or HER-1: e.g. erlotinib, gefitinib, vandetanib, osimertinib, cetuximab, necitumumab, or panitumumab), VEGFR inhibitors or pathway blockers (e.g. bevacizumab, pazopanib, sunitinib, sorafenib, axitinib, regorafenib, ponatinib, cabozantinib, vandetanib, ramucirumab, lenvatinib, ziv-aflibercept), PARP inhibitors (e.g., olaparib, rucaparib, veliparib, or niraparib), JAK inhibitors (JAK1 and/or or JAK2, e.g. ruxolitinib, baricitinib, itacitinib (INCB39110)), LSD1 inhibitors (e.g. INCB59872 and INCB60003), TDO inhibitors, PI3K-delta inhibitors (e.g. INCB50465 and INCB50797), PI3K-gamma selective inhibitors PI3K-gamma inhibitors such as Pim inhibitors (e.g. INCB53914), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl and Mer), adenosine receptor antagonists (e.g. A2a/A2b receptor antagonists ), HPK1 inhibitors, chemokine receptor inhibitors (e.g. CCR2 or CCR5 inhibitors), SHP1/2 phosphatase inhibitors, histone deacetylase inhibitors (HDACs) such as HDAC8 inhibitors, angiogenesis inhibitors, interleukins Included are receptor inhibitors, bromo and extra terminal family member inhibitors (eg bromodomain inhibitors or BET inhibitors such as INCB54329 and INCB57643), or combinations thereof.

In some embodiments, the therapeutic methods described herein are combined with administration of a PI3Kδ inhibitor. In some embodiments, the therapeutic methods described herein are combined with administration of a JAK inhibitor. In some embodiments, the therapeutic methods described herein are combined with administration of a JAK1 or JAK2 inhibitor (eg, baricitinib or ruxolitinib). In some embodiments, the therapeutic methods described herein are combined with administration of a JAK1 inhibitor. In some embodiments, the therapeutic methods described herein are combined with administration of a JAK1 inhibitor that is selective over JAK2.

Exemplary antibodies for use in combination therapy include trastuzumab (eg, anti-HER2), ranibizumab (eg, anti-VEGF-A), bevacizumab (AVASTIN™, eg, anti-VEGF), panitumumab (eg, anti- EGFR), cetuximab (eg, anti-EGFR), Rituxan (eg, anti-CD20), and antibodies directed against c-MET.

One or more of the following agents may be administered to a patient in combination with the treatment methods of the present disclosure, provided as a non-limiting list: cytostatics, cisplatin, doxorubicin, taxotere, taxol, Etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilone, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH66336, R115777, L778,123, BMS214662, IRESSA™ (gefitinib), TARCEVA ( (erlotinib), antibody to EGFR, intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine , lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovirine, ELOXATIN™ (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17. Alpha-ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megstrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimi estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelben, anastrazole, letrazole, capecitabine, reloxafine, droloxa fin, hexamethylmelamine, avastin, HERCEPTIN™ (trastuzumab), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (ibritumomab tiuxetan), TRISENOX™ (three arsenic oxide), XELODA™ (capecitabine), vinorelbine, porfimers, ERBITUX™ (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, lerozole, fulvestrant, exemestane, ifosfomide, rituximab, C225 (cetuximab), Campath (alemtuzumab), clofarabine, cladribine, aphidicolone, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP, and MDL-101,731.

The therapeutic methods and regimens of the disclosure can further be used in combination with other methods of treating cancer, for example, by chemotherapy, radiotherapy, tumor-targeted therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapies include cytokine treatment (eg interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, bispecific or multispecific antibodies, Antibody drug conjugates, adoptive T cell transfer, Toll receptor agonists, RIG-I agonists, oncolytic virotherapy and immunomodulatory small molecules such as thalidomide or JAK1/2 inhibitors, PI3Kdelta inhibitors and the like. A compound may be administered in combination with one or more anti-cancer agents, such as chemotherapeutic agents. Examples of chemotherapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, intravenous busulfan, oral For busulfan, carsterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftit doxorubicin, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alpha 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, Leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone fenpropionate, nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronate, panitumumab , pegasparagase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, test lactones, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and and zoledronate.

Additional examples of chemotherapeutic agents include proteosome inhibitors (eg, bortezomib), thalidomide, Revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

Exemplary steroids include corticosteroids such as dexamethasone or prednisone.

Exemplary Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib, and ponatinib, and pharmaceutically acceptable salts. Other exemplary suitable Bcr-Abl inhibitors include the classes and species disclosed in US Pat. No. 5,521,184, WO04/005281, and US Serial No. 60/578,491, and Pharmaceutically acceptable salts thereof are included.

Exemplary suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linifanib, sunitinib, sunitinib, maleate, sorafenib, quizartinib, crenolanib, pacritinib, tanzutinib, PLX3397 and ASP2215, and pharmaceutically acceptable salts thereof. mentioned. Other exemplary suitable Flt-3 inhibitors include compounds such as those disclosed in WO03/037347, WO03/099771, and WO04/046120, and pharmaceutically acceptable salts thereof.

Exemplary suitable RAF inhibitors include dabrafenib, sorafenib, and vemurafenib, and pharmaceutically acceptable salts thereof. Other exemplary suitable RAF inhibitors include compounds such as those disclosed in WO00/09495 and WO05/028444, and pharmaceutically acceptable salts thereof.

Exemplary suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520, and GSK2256098, and pharmaceutically acceptable salts thereof. Other exemplary suitable FAK inhibitors include compounds as disclosed in WO04/080980, WO04/056786, WO03/024967, WO01/064655, WO00/053595, and WO01/014402, and their pharmaceutical acceptable salts.

Exemplary suitable CDK4/6 inhibitors include palbociclib, ribociclib, trilaciclib, lerosiclib, and abemaciclib, and pharmaceutically acceptable salts thereof. Other exemplary suitable CDK4/6 inhibitors include compounds as disclosed in WO09/085185, WO12/129344, WO11/101409, WO03/062236, WO10/075074, and WO12/061156, and their Pharmaceutically acceptable salts are included.

In some embodiments, compounds of the present disclosure may be used in combination with one or more other kinase inhibitors, including imatinib, particularly to treat patients resistant to imatinib or other kinase inhibitors. .

In some embodiments, the therapeutic methods of the present disclosure can be used in combination with chemotherapeutic agents in the treatment of cancer without exacerbating their toxic effects compared to the response of the chemotherapeutic agents alone. , may improve treatment response. In some embodiments, the treatment methods of the present disclosure can be used in combination with chemotherapeutic agents provided herein. For example, additional pharmaceutical agents used to treat multiple myeloma may include, but are not limited to, melphalan, melphalan plus prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). . Still additional agents used to treat multiple myeloma include Bcr-Abl, Flt-3, RAF and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, proteasome inhibitor, corticosteroid, or immunomodulatory agent. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable results of combining the therapeutic methods of the present disclosure with additional agents.

Agents may be combined with epacadostat and/or human PD-1 or human PD-L1 or antigen-binding fragments thereof in a single or sequential dosage form of the method of treatment, or agents may be administered simultaneously or as separate dosage forms. They can be administered sequentially.

In some embodiments, a corticosteroid such as dexamethasone is administered to a patient in combination with the treatment methods of the present disclosure, where dexamethasone is administered intermittently as opposed to continuously.

The therapeutic methods described herein include cancer cells, purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), cells, and other cells such as cells transfected with genes encoding immunostimulatory cytokines. of immunogenic agents. Non-limiting examples of tumor vaccines that may be used include peptides of melanoma antigens, such as peptides of gp100, MAGE antigen, Trp-2, MRTI and/or tyrosinase, or to express the cytokine GM-CSF. Includes transfected tumor cells.

The therapeutic methods described herein can be used in combination with vaccination protocols for the treatment of cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins derived from viruses involved in human cancer, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposiherpes sarcoma virus (KHSV). mentioned. In some embodiments, the therapeutic methods and regimens of the disclosure may be used in combination with tumor-specific antigens, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, the therapeutic methods described herein can be combined with dendritic cell immunization to activate potent anti-tumor responses.

The therapeutic methods and regimens of the present disclosure can be used in combination with bispecific macrocyclic peptides that target Fealpha or Fegamma receptor-expressing effector cells to tumor cells. The therapeutic methods and regimens of the present disclosure can also be combined with macrocyclic peptides that activate host immune responsiveness.

In some further embodiments, the therapeutic methods of the present disclosure can be combined with other therapeutic agents and administered to the patient before, during, and/or after bone marrow or stem cell transplantation. The therapeutic methods and regimens of the present disclosure can be used in combination with bone marrow transplantation for the treatment of various tumors of hematopoietic origin.

When two or more pharmaceutical agents are administered to a patient, they can be administered simultaneously, separately, sequentially, or in combination (e.g., three above drugs).

Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in standard literature. For example, many administrations of chemotherapeutic agents are described in the "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is given in its entirety. is incorporated herein by reference as well.

II. Immune Checkpoint Therapy The therapeutic methods of the present disclosure can be used in combination with administration of one or more immune checkpoint inhibitors or agonists (eg, antibodies or small molecules) for the treatment of diseases such as cancer. Exemplary immune checkpoint inhibitors include CBL-B, CD20, CD28, CD40, CD70, CD122, CD96, CD73, CD47, CDK2, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, Arginase, HPK1 , CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TLR (TLR7/8), TIGIT, CD112R, VISTA. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137 (4-1BB). In some embodiments, the compounds provided herein are one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFR beta inhibitors can be used in combination with

In some embodiments, inhibitors of immune checkpoint molecules are inhibitors of KIR, TIGIT, LAIR1, CD160, 2B4 and TGFR beta.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CTLA-4, eg, an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

In some embodiments, the inhibitor is MCLA-145.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of LAG3, eg, an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, INCAGN2385, or eftiragimod alfa (IMP321).

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is olelumab.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of TIGIT. In some embodiments, the inhibitor of TIGIT is OMP-31M32.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of VISTA. In some embodiments, the inhibitor of VISTA is JNJ-61610588 or CA-170.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of B7-H3. In some embodiments, the inhibitor of B7-H3 is enobrituzumab, MGD009, or 8H9.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of KIR. In some embodiments, the inhibitor of KIR is lililumab or IPH4102.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of A2aR. In some embodiments, the inhibitor of A2aR is CPI-444.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of TGF-beta. In some embodiments, the inhibitor of TGF-beta is Travedersen, Garsertinib, or M7824.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of PI3K-gamma. In some embodiments, the inhibitor of PI3K-gamma is IPI-549.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CD47. In some embodiments, the inhibitor of CD47 is Hu5F9-G4 or TTI-621.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is MEDI9447.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CD70. In some embodiments, the inhibitor of CD70 is cusatuzumab or BMS-936561.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of TIM3, eg, an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

In some embodiments, the inhibitor of immune checkpoint molecules is an inhibitor of CD20, eg, an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

In some embodiments, the immune checkpoint molecule agonist is an agonist of OX40, CD27, CD28, GITR, ICOS, CD40, TLR7/8, and CD137 (also known as 4-1BB).

In some embodiments, the agonist of CD137 is urelumab. In some embodiments, the agonist of CD137 is utomilumab.

In some embodiments, the immune checkpoint molecule agonist is a GITR agonist. In some embodiments, the GITR agonist is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, MEDI1873, or MEDI6469.

In some embodiments, the immune checkpoint molecule agonist is an OX40 agonist, eg, an OX40 agonist antibody or an OX40L fusion protein. In some embodiments, the OX40 agonist antibody is INCAGN01949, MEDI0562 (tavolimab), MOXR-0916, PF-04518600, GSK3174998, BMS-986178, or 9B12. In some embodiments, the agonist of the OX40L fusion protein is MEDI6383.

In some embodiments, the immune checkpoint molecule agonist is an agonist of CD40. In some embodiments, the agonist of CD40 is CP-870893, ADC-1013, CDX-1140, SEA-CD40, RO7009789, JNJ-64457107, APX-005M, or Chi Lob 7/4.

In some embodiments, the immune checkpoint molecule agonist is an ICOS agonist. In some embodiments, the ICOS agonist is GSK-3359609, JTX-2011, or MEDI-570.

In some embodiments, the immune checkpoint molecule agonist is an agonist of CD28. In some embodiments, the CD28 agonist is ceralizumab.

In some embodiments, the immune checkpoint molecule agonist is an agonist of CD27. In some embodiments, the CD27 agonist is vallilumab.

In some embodiments, the immune checkpoint molecule agonist is a TLR7/8 agonist. In some embodiments, the TLR7/8 agonist is MEDI9197.

The therapeutic methods and regimens of the disclosure may be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody comprises PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3 or TGFβ receptor. target. In some embodiments, bispecific antibodies bind to PD-1 and PD-L1. In some embodiments, the bispecific antibody that binds PD-1 and PD-L1 is MCLA-136. In some embodiments, bispecific antibodies bind to PD-L1 and CTLA-4. In some embodiments, the bispecific antibody that binds PD-L1 and CTLA-4 is AK104.

In some embodiments, compounds of the present disclosure may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an inhibitor of TDO or arginase.

As provided throughout, additional compounds, inhibitors, agents, etc. may be combined with the present compounds in a single or sequential dosage form, or they may be administered simultaneously or sequentially as separate dosage forms. can be

Labeled Compounds Another aspect of the present disclosure is that the present disclosure will be useful in both in vitro and in vivo assays, as well as imaging techniques, to localize and quantify IDO1 in tissue samples, including humans. It relates to labeled compounds (radiolabels, fluorescent labels, isotope labels, etc.) of the disclosure.

The disclosure further includes isotopically labeled epacadostat. An "isotopically" or "radiolabeled" compound has one or more atoms having an atomic mass or mass number different from those typically found in nature (i.e., naturally occurring) Epacadostat of the present disclosure replaced or substituted by an atom having Suitable radionuclides that can be incorporated into compounds of the present disclosure include ² H (also written as D for deuterium), ³ H (also written as T for tritium), ¹¹ C, ¹³ C, ¹⁴ C , ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³⁵ S, ³⁶ Cl, ⁸² Br, ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, ¹²³ I, ¹²⁴ I, ¹²⁵ I, and ¹³¹ I including but not limited to; For example, one or more hydrogen atoms in the compounds of the present disclosure can be replaced with deuterium atoms and optionally replaced with deuterium atoms.

One or more constituent atoms of epacadostat may be substituted with isotopes of atoms, in natural or unnatural abundance, or may be substituted. In some embodiments, epacadostat contains at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of this disclosure can be replaced or replaced with deuterium. In some embodiments, the compounds contain 2 or more deuterium atoms. In some embodiments, the compound contains 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all hydrogen atoms in a compound can be or can be replaced by deuterium atoms.

Synthetic methods for incorporating isotopes into organic compounds are known in the art (New York, NY, Appleton-Century-Crofts, 1971, The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau 2007, 7744-7765, The Organic Chemistry of Isotopic Labeling by James R. Hanson, Royal Society of Chemistry 2). Isotopically-labeled compounds can be used in a variety of studies, such as NMR spectroscopy, metabolic studies, and/or assays.

Substitution with heavier isotopes, such as deuterium, may provide certain therapeutic advantages due to higher metabolic stability, e.g., increased in vivo half-life or decreased dosing regimens, and thus in some circumstances may be preferred (see for example A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210, R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312 In particular, substitutions at one or more sites of metabolism may confer one or more therapeutic benefits.

A "radiolabel" or "labeled compound" is understood to be a compound that incorporates at least one radionuclide. In some embodiments, the radionuclide is selected from the group consisting of ^3H and ^14C . In some embodiments, the radionuclide is selected from the group consisting of ¹¹ C, ¹⁸ F, ⁷⁵ Br, ⁷⁶ Br, and ⁷⁷ Br.

Kits The present disclosure also includes pharmaceutical kits useful, for example, for the treatment of the cancers and solid tumors referred to herein, which contain one of the pharmaceutical compositions described herein. Including the above containers. As will be appreciated by those skilled in the art, such kits may be configured in a variety of conventional pharmaceutical kit configurations, e.g., containers with one or more pharmaceutically acceptable carriers, additional containers, etc., if desired. It can further include one or more of the elements. Instructions, either as an insert or a label, indicating quantities of ingredients to be administered, guidelines for administration, and/or guidelines for mixing the ingredients can also be included in the kit.

Following are examples of the practice of the invention. They should in no way be construed as limiting the scope of the invention.

The following examples are provided to better illustrate the claimed invention and are not intended to limit the scope of the invention. To the extent a particular material is mentioned, it is for illustrative purposes only and is not intended to limit the invention. Those skilled in the art may develop equivalent means or reactants without exercising the power of this invention and without departing from its scope.

Example 1. Phase 1b Trial of Epacadostat in Combination with Antibody X General Study Design This trial is an open-label, non-randomized, multicenter, Phase 1b trial with independent treatment arms. This study consists of two parts. 1) dose escalation to find the maximum tolerated dose (MTD)/recommended phase 2 dose (RP2D) of the combination of Antibody X and epacadostat, and 2) escalation at the selected dose to assess safety and clinical Further investigation of preliminary evidence of activity.

Dose escalation uses a Bayesian optimal interval (BOIN) design with a cohort size of approximately 3 evaluable participants. A target rate of dose-limiting toxicity (DLT) is assumed to be 30% for each combination. Up to 9 participants will be enrolled at each dose level. Dose levels for combinations of Antibody X and epacadostat are shown in Table 1.

Treatment arms will be enrolled in parallel in a non-randomized manner with participants assigned to the open cohort by the sponsor or designee. Prioritize open dose escalation cohorts. If more than one dose expansion cohort is available, participants will be alternately assigned until enrollment is completed, taking into account available data on combinations in the participant's tumor type. Based on new pharmacokinetic (PK) or pharmacodynamic data (including exploratory immunoassay results), additional dose levels or schedules may be explored, or some of the dose escalation cohorts may be expanded It does not have to be open. Intermediate dose levels or alternative dose schedules can be explored to collect additional safety, PK, and pharmacodynamic data. Also, if the higher dose level exceeds the MTD, intermediate dose levels may be explored.

Participants in the dose escalation cohort will be observed for the occurrence of DLT for 28 days. Participants receiving the combination of Antibody X and epacadostat must receive at least 75% of the oral dose evaluable for DLT.

Once the combination RP2D is determined, continuing participants receiving the lower doses will tolerate the current dose if the participant meets protocol eligibility criteria at escalation and has no drug-related toxicity grade 2 or greater, If the investigator determines that a participant could potentially benefit from a higher dose, titration to RP2D may be permitted with medical monitor approval.

MTD is defined as the highest dose at which less than about one-third of participants have a DLT. Dose-limiting toxicities occurred during the first 28 days of treatment, leading to dose escalation and determination of MTD and RP2D. In addition, participants with a late safety event meeting the definition of DLT or with intolerable persistent toxicity determined to be attributable to any of the study drugs (e.g., grade 2 peripheral nerve obstacles) are considered in the selection of each combination RP2D. RP2D can be selected from any of the available dose levels that do not exceed the MTD. If the MTD is not reached, RP2D is selected from available doses based on safety, pharmacokinetic (PK), and translational data.

A baseline tumor biopsy sample will be obtained from all participants. The treatment cycle is 28 days unless otherwise stated. At the start of each treatment cycle after Cycle 1, participants must meet the following criteria.
(i) hemoglobin ≧8 g/dL
(ii) ANC≧1.0×10 ⁹ /L
(iii) platelet count ≧75×10 ⁹ /L
(iv) ALT/AST/bilirubin < grade 2
(v) Resolution of all immune-related therapy emerging adverse events (TEAEs) to grade 1 or less (except hyperglycemia [allowed to grade 2] and endocrine disorders controlled by hormone replacement);
(vi) Resolution of all non-immune related TEAEs to grade 1 or less or baseline (except grade 2 alopecia). A transient asymptomatic laboratory elevation <Grade 3 does not require dose interruption if the participant is asymptomatic, the elevation is not clinically significant, and is discussed with the medical monitor.

The duration of treatment in trials is a maximum of 2 years in the absence of clinical progression or intolerable toxicity. The trial will end after the last participant in each treatment group has been followed for approximately 6 months.

Participants are eligible to participate in this study only if all of the following criteria apply.
• Ability to understand and demonstrate willingness to sign a written ICF for the clinical trial.
● Adult men and women over the age of 18 (or depending on local country requirements).
- Participants with histologically proven, unresectable or metastatic locally advanced solid tumors for whom no approved therapy with demonstrated clinical benefit is available or against standard therapy participants who were intolerant or refused.
• Tumor lesions measurable or non-measurable according to RECIST v1.1. (Note: Participants enrolled in the dose escalation cohort must have at least one lesion that can be biopsied).
• Willingness to provide fresh or archival tumor tissue for correlative studies.
Eastern Cooperative Oncology Group (ECOG) Performance Status 0-1
• Willingness to avoid conceiving or fathering children based on certain criteria.

Participants will be excluded from the study if they meet any of the following criteria:
• Receive chemotherapy within 21 days of the first dose of study treatment, excluding local radiation therapy.
• Toxicity of prior therapy not grade 1 or less or not reversing to baseline (except alopecia and anemia not requiring infusion support).
• Participants with clinical laboratory values at screening as defined in Table 2.
• Active autoimmune diseases that require systemic immunosuppression that exceeds physiological maintenance doses of corticosteroids.
• Known active CNS and/or leptomeningeal metastatic carcinoma.
Known additional malignancies that are ongoing or require aggressive treatment, or history of other malignancies within 2 years of study entry (cured basal or squamous cell carcinoma of the skin, superficial bladder cancer, Prostate intraepithelial neoplasia, cervical carcinoma in situ, or other non-invasive or indolent malignancies, or cancers for which participants were disease-free for >1 year after treatment with curative intent).
- Known active hepatitis A, B, or C defined by elevated transaminases with the following serological characteristics: hepatitis A virus IgM antibodies without previous immunization, anti-hepatitis C virus , anti-hepatitis B core antigen IgG or IgM, or positivity for hepatitis B surface antigen.
• Active infections requiring systemic antibiotics.
• Grade 2 or greater immune-related toxicity while receiving prior immunotherapy.
• Known hypersensitivity to either the investigational drug, excipients, or another monoclonal antibody that cannot be controlled by standard means (eg, antihistamines and corticosteroids).
- Participants with cardiac dysfunction or clinically significant heart disease:
o New York Heart Association Class III or IV heart disease, including pre-existing clinically significant ventricular arrhythmia, congestive heart failure, or cardiomyopathy o Unstable angina pectoris within 6 months prior to study entry .
o Acute myocardial infarction within 6 months prior to study entry.
o Other clinically significant cardiac disease (i.e., grade 3 or greater hypertension, history of unstable hypertension, or poor compliance with antihypertensive regimens) from prior treatment-related toxicities (baseline or grade 1 or less) must be recovered.
●Women who are breastfeeding.
• If the subject underwent major surgery, he or she must have fully recovered from interventional toxicities and/or complications prior to initiation of study treatment.
- Have received a live vaccine within 30 days of the start of planned investigational treatment.
- Evidence of interstitial lung disease or active non-infectious pneumonia.
- Current use of prohibited substances, including other anticancer therapies, including investigational therapies, immunosuppression above physiological maintenance corticosteroid doses (except for acute treatment of AEs), leukocyte transfusions, investigational and 5 half-lives Live vaccines, products containing more than a total daily dose of 2 g or 2000 mg of acetyl-para-aminophenol, MAOIs or drugs associated with significant MAO inhibitory activity, the final dose of epacadostat should be discontinued 21 days prior to initiation of study treatment. Prohibited until 14 days after taking. Coumarin anticoagulant.
Any condition that, at the investigator's discretion, prevents full participation in the study, poses a significant risk to the participant, or interferes with the interpretation of study data, including administration of study medication and attendance at required study visits .
• Participants must not have a history of serotonin syndrome after being administered one or more serotonergic drugs.
Participants known to be HIV-positive, unless they meet all of the following criteria:
o CD4+ counts ≧300/μL.
o Undetectable viral load.
o Receiving very active antiretroviral therapy.
• Participants must not have a history of gastrointestinal disease (eg, inflammatory bowel disease, Crohn's disease, ulcerative colitis) that may affect drug absorption.

Table 3 shows the Investigational Treatment Information for Infused and Oral Investigational Drugs. At visits where oral study drug is administered at the clinic, oral study drug is administered immediately prior to the start of the Antibody X infusion.

Dose-Limiting Toxicities A DLT is defined as the occurrence of any of the toxicities listed in Table 4, which are probable, probable, or certain to follow study treatment from the start of treatment up to and including Day 28. This is due to All DLTs will be assessed by the investigator using the Common Terminology Criteria for Adverse Events: Version 5 (CTCAE v5) criteria. Participants receiving Antibody X in combination with oral study drug must receive at least 75% of the oral dose to be evaluable for DLT. If study treatment is discontinued due to drug-related toxicity, this is considered a DLT.

At the start of each treatment cycle, participants must meet the above treatment continuation criteria prior to Antibody X infusion. If criteria are not met, study treatment (both study drugs) will be discontinued. Participants will be removed from the active treatment portion of the trial if the treatment continuation criteria are not met within 28 days of the scheduled cycle start date. If any study drug combination must be discontinued due to unacceptable toxicity, the subject will be withdrawn from both study drugs (ie, study treatment) and entered the follow-up portion of the study.

Dose reductions are not permitted for Antibody X and epacadostat.

Response Assessment Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 guidelines will be followed to assess response in solid tumors. The recommended method for measuring and tracking tumor burden is determined by CT scans performed using consistent techniques and facilities. Alternative methods of CT scanning (eg, MRI) can be substituted at the investigator's discretion. provided that the same method was used throughout the study and that the method was consistent with RECIST v1.1. Initial tumor imaging will be performed within 28 days of the first dose of study treatment. Tumor lesions located in previously irradiated areas or areas that received other local therapy are not selected as target lesions. Additionally, it is recommended that tumor lesions selected for biopsy not be selected as target lesions.

Immunotherapeutic agents can produce anti-tumor effects by enhancing endogenous cancer-specific immune responses. The pattern of response seen with such approaches may extend beyond the typical time course of responses seen with cytotoxic agents, leading to an initial increase in tumor burden, or the appearance of new lesions followed by clinical responses. can be shown. Standard RECIST v1.1 may not provide a completely accurate response assessment for immunotherapeutic agents, excluding participants from treatment who might otherwise have benefited from further immunotherapy treatment may be required. Therefore, the general principles of a modified version of RECIST v1.1 for immune-based therapeutics, called iRECIST, are used in the evaluation of participant responses in the exploratory capacity of this study. The use of iRECIST includes a requirement for confirmation of progression to describe immunotherapy response patterns and to rule out or confirm pseudoprogression.

Adverse events will be monitored with all serious adverse events (SAEs) recorded and reported. Laboratory tests are performed, including measurement of kynurenine levels in plasma and tumor samples.

World Wide Clinical Trials, Inc.; , plasma kynurenine levels were measured by an LC-MS/MS method. Patient samples were obtained pre-dose and at defined times post-treatment. Plasma kynurenine levels were measured according to Huang, et al. , Bioanalysis, 2013;5(11):1397-1407.

Kynurenine levels in flash-frozen tumor samples are measured by quantitative mass spectrometry imaging or LC-MS/MS. Tumor biopsies are obtained before treatment and during the fifth week of treatment.

Follow-up Analyzes Participants who discontinued study treatment for reasons other than disease progression should be transferred to the disease status follow-up period and evaluated by radiographic imaging every 12 weeks ± 7 days to monitor disease status. be. Efforts can be made to collect information about disease status until initiation of new anticancer therapy, disease progression, death, study termination, and loss of follow-up participants. Once the participant has received the last dose of study therapy, confirmed disease progression, or initiated a new anticancer drug treatment, the participant enters the Survival Follow-up Period, with telephone calls at least every 12 weeks, Patients must be contacted by e-mail or in-office to assess survival to death, withdrawal of consent, or end of study.

Results In the above study, two groups (groups 1 and 2) of 3 patients with solid tumors were given Antibody X (500 mg Q4W) in combination with 100 mg BID or 600 mg BID epacadostat. rice field. Patients in Group 1 received 100 mg BID epacadostat and Group 2 received 600 mg BID epacadostat. Patients in Group 2 showed increased reduction in plasma kynurenine levels compared to 3 patients in Group 1 at day 8 of treatment, with 2/3 sustained reductions after 5 weeks of treatment showed that. This suggests that higher doses of epacadostat result in higher levels of IDO1 inhibition.

This reduction in plasma kynurenine levels with 600 mg BID epacadostat is surprising based on the results of previous clinical trials using 300 mg BID epacadostat in combination with another anti-PD-1 antibody, pembrolizumab (200 mg/kg Q3W). , I _max , I _min and I _avg values were 97%, 76% and 88%.

The term "I _max " refers to the maximum percentage of IDO inhibition calculated over all PK time points. I _max is the maximum or maximum percentage of IDO inhibition during which drug is administered to that trough (eg, the lowest concentration of drug present in a subject). For example, for twice daily dosing, I _max refers to the maximum percentage of IDO inhibition between 0 hours (before dosing) and 12 hours after dosing.

The term "I _min " refers to the minimum percentage of IDO inhibition calculated over all PK time points. I _min is the percentage of IDO inhibition in the trough (eg, generally at 12 hours for twice daily dosing). For example, I _min ≧50 refers to 50% or greater IDO inhibition in the trough (eg, at 12 hours).

The term “I _avg ” refers to the average percentage of IDO inhibition during the period in which the drug is administered trough. Calculated as the area under the inhibition curve (AUC) over time (calculated using the linear trapezoidal method) divided by the dosing interval (eg, 12 hours for BID dosing).

Calculated I _max , I _min and I _avg values for each subject were summarized as the mean±standard deviation (geometric mean) standard statistical calculation for each dose group.

The combination of Antibody X and epacadostat is being evaluated in a dose-finding study (INCMGA0012-102, NCT03059823). Thirty-one participants were treated with a combination of epacadostat at doses of Antibody X 500 mg Q4W and 100 mg, 400 mg, 600 mg, and 900 mg BID. Epacadostat 900 mg BID exceeded the MTD based on the development of grade 3 rash in 2 of 3 participants, with the 3rd participant developing rash shortly after the protocol-defined DLT window. Treatment-on-treatment adverse events (TEAEs) reported in 10% or more of the participants included fatigue, nausea, abdominal pain, itching, maculopapular rash, and diarrhea. Serious adverse events (SAEs) occurred in 8 participants (25.8%), but no SAEs occurred in >1 participant. Three participants had dose-limiting toxicities (DLTs), all of which were grade 3 maculopapular rashes (1 DLT occurred at the 400 mg BID dose of epacadostat in combination with Antibody X; man occurred on the 900 mg BID dose of epacadostat). Epacadostat 600 mg BID was well tolerated in combination with Antibody X 500 mg Q4W in an initial cohort of participants and is being further evaluated. In addition, epacadostat 600 mg BID produced sustained normalization of kynurenine in preliminary observations.

FIG. 1 shows plasma kynurenine results for patients treated with Antibody X in combination with the indicated doses of epacadostat (100 mg BID, 400 mg BID, 600 mg BID, 900 mg BID). Plasma kynurenine was measured before treatment (C1D1) and at the indicated visits. FIG. 1 shows that treatment with 600 mg BID resulted in a sustained (up to 4 months) reduction in plasma kyn in most patients.

Example 2. Phase 2 Clinical Trial of Antibody X in Combination with Epacadostat in Patients With Recurrent or Progressive PD-L1 Positive Microsatellite-stable Endometrial Cancer General Study Design This is microsatellite stable (MSS) and PD-L1 positive. A multicenter, non-labeled, non-randomised phase 2 trial of Antibody X in combination with epacadostat in participants with advanced or metastatic endometrial cancer that had progressed during or after platinum-based chemotherapy. Participants will receive Antibody X 500 mg Q4W (administered IV) combined with Epacadostat 600 mg BID (administered PO) for up to 26 cycles. The trial will include one interim analysis for futility after 24 participants have been enrolled. Table 5 shows the objectives and endpoints of this study.

After discontinuation of study treatment, the treatment portion of the study ends and subjects enter follow-up. Follow-up consists of three parts: safety follow-up, disease status follow-up, and survival follow-up. Participants will be followed for safety for 90 days after the last dose of study treatment or until initiation of new anticancer therapy, whichever comes first. Participants who discontinued study treatment for reasons other than disease progression will be moved to a follow-up period for disease status, where Q8W will continue to be assessed to determine whether new anticancer therapy is started, disease progression, death, study treatment Disease status should be monitored until termination or until the participant is lost from follow-up.

BACKGROUND AND RATIONALE Blockade of immune-inhibitory pathways is emerging as an important therapeutic modality for the treatment of cancer, as evidenced by the clinical responses observed with antibodies against PD-1/PD-L1. Although these single agents have antitumor activity, multiple immune-inhibitory mechanisms exist simultaneously within the tumor microenvironment, suggesting that combination therapy may be required for optimal therapeutic efficacy. (Quezada & Peggs, Br. J. Cancer. 2013, 108: 1560-1565). The aim of this study was to demonstrate that Antibody X, a PD-1 inhibitor, and epacadostat, an IDO1 inhibitor, may improve the therapeutic efficacy of anti-PD-1 monotherapy in patients with PD-L1-positive, MSS endometrial cancer. To investigate the safety and efficacy of the combination of

Endometrial cancer (EC) is the most common gynecologic cancer in developed countries (Colombo et al, Int. J. Gynecol. Cancer 2016, 26:2-30). In 2018, it is estimated that approximately 380,000 new cases of endometrial cancer were diagnosed worldwide and 90,000 women worldwide died from the disease. Endometrial cancer is the sixth most common cancer in women worldwide (Brey et al, CA Cancer J. Clin. 2018, 68:394-424). In 2020, approximately 65,620 new cases and 12,590 deaths from endometrial cancer are expected in the United States. Two-thirds of new cases are diagnosed early. Mean age at presentation is 60 years and is rare in women younger than 45 years. The incidence of endometrial cancer has increased over time and successive generations in many countries around the world, especially in countries with rapid socioeconomic transitions (Lortet-Tieulent et al, J. Med. Natl. Cancer Inst. 2018, 110:354-361). Although the 5-year survival rate for localized disease is 95%, only 17% of women with distant metastatic disease are expected to live 5 years after diagnosis.

Risk factors for endometrial cancer include elevated estrogen levels (caused by obesity, diabetes, and a high-fat diet), menarche at early age, impotence, menopause at late age, advanced age (≥55 years), and the use of tamoxifen (Van den Bosch et al, Best Pract. Res. Clin. Obstet. Gynaecol. 2012, 26:257-66, Kitchener & Trimble, Int. J. Gynecol. Cancer, 2009, 19:134-140 2013, 2013:583891, Obermair et al, Int. J. Cancer, 2010, Dec 1, 127:2678-2684). Obesity with a BMI greater than 30 is responsible for up to 81% of newly diagnosed endometrial cancers (Nevadunsky et al, Obstet. Gynecol. 2014, 124:300-306). The incidence of endometrial cancer is increasing mainly due to the increased incidence of obesity and consequent hyperinsulinemia.

Although most endometrial cancers are sporadic, 2-5% of cases are familial and harbor germline mutations in mismatch repair genes (Lynch et al, Nat. Rev. Cancer, 2015, 15: 181-194). Four molecular clusters of ECs have been identified in a comprehensive study of 373 ECs through The Cancer Genome Atlas (TCGA) (Kandoth et al, Nature, 2013, 497:67-73). These are: (1) hypermutation/polymerase ε (POLE) mutation, (2) hypermutation/MSI (MSI-H), (3) low copy number (microsatellite stable [MSS]), and (4) high copy number. POLE tumors had the best PFS and high copy number tumors the worst. Unfortunately, the genome sequencing method used by TCGA is not suitable for wider clinical application. Localized endometrial cancer can be cured by surgical resection. Systemic therapy is used in more advanced disease. Hormone therapy is preferred in low-grade, hormone-positive disease that does not progress rapidly. Not recommended for patients with visceral disease and rapidly progressive disease (Colombo et al, Int. J. Gynecol. Cancer, 2016, 26:2-30). Endometrial cancer is chemosensitive, and multi-drug chemotherapy is preferred for metastatic, recurrent, or high-risk disease (Colombo et al, Int. J. Gynecol. Cancer, 2016, 26:2-30 , National Comprehensive Care Network. Clinical Practice Guidelines in Oncology. Uterine Neoplasms. Version 3.2019-11 February 2019). Anthracyclines, taxanes, and platinum-based compounds have been extensively investigated in this disease. A combination of carboplatin and paclitaxel is commonly used as first-line therapy in advanced EC, with an ORR of approximately 50%, a PFS of 13 months, and an OS of 3 years (Miller et al., Gynecol. Oncol. 2012, 125:771-773, Colombo et al., Int. J. Gynecol. Cancer, 2016, 26:2-30).

Treatment options after failure of first-line chemotherapy are limited (Fleming et al, J. Clin. Oncol. 2015, 33:3535-3540). After failure of first-line chemotherapy, there are no established effective second-line agents for this disease. Paclitaxel had the highest RR of 25% in patients previously treated with cisplatinum and doxorubicin. Patients receiving paclitaxel as first-line therapy have only an 8% RR with docetaxel. The 5-year survival rate for advanced/recurrent measurable disease after second-line therapy is <10% (Moxley et al, The Oncologist, 2010, 15:1026-1033, Dizon et al, J. Clin. Oncol.2009, 27:3104-3108 and Garcia et al, Gynecol.Oncol.2008, 111:22-26). Everolimus plus letrozole and bevacizumab have also shown moderate activity in small uncontrolled trials, as well as PD-1 inhibitor monotherapy and in combination with other therapies in tumors not selected for abnormal DNA repair. (Ott et al, J. Immunother. Cancer 2017, 5:16 and Oaknin et al, Gynecol. Oncol. 2019, 154(1 suppl): Abstract 33). In particular, MMR deficiency has been associated with resistance to commonly used chemotherapeutic agents (Guillotin & Martin, Exper. Cell Res. 2014, 329:110-115). In approximately 25-30% of ECs, tumors are MMR-deficient or MSI-H (Murali et al, Lancet Oncol. 2014, Jun; 15(7):e268-278, Karamurzin and Rutgers, Int. J. Gynecol .Pathol.2009, 28:239-255). Promising clinical activity using immunotherapy-based approaches has been seen in tumors characterized by DNA repair defects (e.g., MSI-H, dMMR, or POLE hypermutation) associated with high neoantigen load (Mittica et al. et al, Oncotarget, 2017, 8: 90532-90544, Brooks et al, CA Cancer J. Clin. 2019, 69: 258-279 and Di Tucci et al, J. Gynecol. Oncol. 2019, 30: e46). Pembrolizumab has been shown to be effective in treating MMR-deficient tumors, including MMR-deficient endometrial cancer (Le et al, N. Engl. J. Med. 2015, 372:2509-2520). In the US, it is approved for the treatment of MSI-H or MMR-deficient endometrial cancer that has progressed on previous therapy. The EC ORR was 36% and the duration of response ranged from 4 to 17 months.

However, the majority of ECs contain MSS tumors. There is an unmet need for more effective treatment of MSS endometrial cancer that has progressed after initial platinum-based chemotherapy. EC cells overexpress PD-1 and PD-L1 in 25-75% of cases, the highest among all gynecologic cancers (Herzog et al, Gynecol. Oncol. 2015, 137:204-205). . Monotherapy with anti-PD-(L)-1 antibodies against MSS tumors with intact DNA repair has moderate clinical activity and no survival benefit has been established (Ott et al, J. Immunother. Cancer, 2017, 5:16, Marcus et al, Clin.Cancer Res.2019, 25:3753-3758, and Fleming et al, J. Clin.Onc.2017, 35(15 suppl): Abstract 5585.

Combination therapy with anti-PD-1 antibodies may be more effective. Recently, the combination of pembrolizumab and lenvatinib showed additional benefit in MSI-H and MSS tumors after progression of previous systemic therapy, with an overall response rate of 63.6% at 24 weeks in MSI-H tumors. and 36.2% in participants with MSS tumors (Makker et al, J. Clin. Oncol. 2020; DOI: 10.1200/JCO.19.02627). Grade 3 or 4 adverse events were reported in 66.9% of subjects, and 21% discontinued treatment secondary to adverse events. More combination regimens need to be evaluated in this population to improve the safety and efficacy of currently available therapies.

Furthermore, endometrial cancers have been shown to have much higher amounts of indoleamine-2,3-dioxygenase (IDO) in inflamed tissue compared to tryptophan-2,3-dioxygenase (TDO). It is IDO and TDO are two major enzymes that regulate the initial and rate-limiting steps of the kynurenine pathway. As noted above, local depletion of tryptophan and accumulation of pro-apoptotic kynurenines can profoundly affect T-cell proliferation and survival. Therefore, cancers that express much higher amounts of IDO compared to TDO may respond better to treatment with an IDO inhibitor such as Antibody X and a PD-1 antibody. Current translational datasets show that endometrial cancer expresses 40-fold higher levels of IDO compared to TDO and 60% IDOhi/TDOlow and is more sensitive to treatment with IDO inhibitors such as epacadostat. show. Other cancers with high ratios of IDO:TDO include cervical cancer (IDO:TDO 79:1 and 60% IDOhi/TDOlow), kidney cancer (or kidney clear cell carcinoma (KIRC) (IDO:TDO 45:1 and 60%). %IDOhi/TDOlow), lung cancer including lung adenocarcinoma (IDO:TDO7.5:1 and 25>%IDOhi/TDOlow), head and neck cancer (head and neck squamous cell carcinoma) (IDO:TDO8:1 and 20%IDOhi /TDOlow).As Example 1 shows that higher doses of epacadostat (up to 600 mg) produce sustained (up to 4 months) reductions in plasma kynurenine levels in most patients, these are Cancers should be more responsive to treatment with epacadostat than cancers with low levels of IDO compared to TPO.

Inclusion Criteria Participants are eligible to participate in this study only if all of the following criteria apply.
• Ability to understand and demonstrate willingness to sign a written ICF for the clinical trial.
● Women over the age of 18 (or depending on local country requirements).
- Histologically confirmed diagnosis of advanced or metastatic endometrial cancer (excluding carcinoma and sarcomas of the uterus).
• Radiological evidence of disease progression after treatment with no more than one platinum-containing regimen for advanced or metastatic disease.
o One neoadjuvant/adjuvant chemotherapy in early stages of disease is allowed. Participants may receive a total of up to two platinum-based chemotherapy regimens, as long as one regimen is given in the neoadjuvant or adjuvant treatment setting. Prior hormonal therapy is acceptable in all disease settings.
• Willingness to provide tumor tissue samples (fresh or archived). Tumor tissue is tested intensively for MSS and PD-L1 status.
o Tumors must be PD-L1 positive and MSS for study entry, as defined by central laboratory results.
• Must have at least one measurable tumor lesion per RECIST v1.1.
ECOG performance status 0 or 1.
• Willingness to avoid pregnancy based on the following criteria.
o Females of childbearing potential must have a negative serum pregnancy test at Screening and, from Screening, take appropriate precautions (at least 99%) to avoid pregnancy until 6 months after the last dose of study treatment. (with certainty). Participants should be informed and understood of an approved method that is at least 99% effective in preventing pregnancy.
• Women of non-fertile potential (ie, surgically sterile by hysterectomy and/or bilateral oophorectomy, or amenorrhea for ≥12 months and at least 50 years of age) are eligible.

Study Treatment Information Table 6 lists study treatment information. For clinic-administered visits, epacadostat should be administered immediately prior to the start of the Antibody X infusion. Dose modifications of Antibody X and epacadostat are not permitted. Antibody X will be resumed at 500 mg Q4W if a dose interruption is required to manage a drug-related TEAE.

Example 3. Phase 2/3 Trial of Retifan Limab + Epacadostat vs. Retifan Limab + Placebo in Participants With High-Risk BCG Non-Responsive Non-Muscle Invasive Bladder Cancer General Study Design , a randomized, double-blind, placebo-controlled Phase 2/3 trial in participants with BCG-nonresponsive, high-risk, non-muscle-invasive bladder cancer (NMIBC), according to good clinical practice Carcinoma in situ (CIS) with or without papillary tumors ineligible for or electing to undergo cystectomy performed. Participants will be stratified by PD-L1 status (PD-L1 positive vs. PD-L1 negative) and papillary disease status (papillary disease present at baseline vs. no papillary disease). The study consists of two treatment arms:
Arm A: retifanlimab 500 mg Q4W + placebo BID
Arm B: retifanlimab 500 mg Q4W + epacadostat 600 mg BID

This study consists of two phases. Phase 2 begins with a 2:1 randomization of participants to receive retifanlimab versus placebo or retifanlimab versus epacadostat, respectively. After 150 participants have been enrolled, enrollment will be suspended for participants who will be monitored for response to treatment for up to 6 months. If the end-of-Phase 2 analysis meets the desired criteria, the trial will enroll an additional 150 participants in a 1:2 ratio to receive retifanlimab plus placebo or retifanlimab plus epacadostat, respectively. Randomization will initiate Phase 3 enrollment.

After discontinuation of study treatment, the treatment portion of the study ends and subjects enter follow-up. Follow-up consists of two parts: safety follow-up and disease status follow-up. Safety will be followed for 90 days after the last dose of study treatment or until initiation of new anticancer therapy, whichever comes first. Participants who discontinued study treatment for reasons other than disease progression entered the disease status follow-up period, where new anticancer therapy was started, disease progression, death, study termination, or the participant was not followed up. It should continue to be assessed by Q12W efficacy assessments and monitor disease status until lost from .

Inclusion Criteria Participants are eligible to participate in this study only if all of the following criteria apply.
1. Ability to understand and demonstrate willingness to sign a written ICF for a clinical trial.
2. Men and women over the age of 18 (or according to local country requirements).
3. Pathologically confirmed high-risk NMIBC is defined as carcinoma in situ (CIS) with or without papillary tumor (high grade Ta or T1),
o Predominant histologic component (>50%) must be urothelial (transitional cell) carcinoma 4. Demonstrated insensitivity to BCG (according to February 2018 FDA guidance)
o High-risk NMIBC unresponsive to BCG is defined as: persistent or recurrent CIS alone or recurrent Ta/T1 (papillary disease/tumor in situ) within 12 months of completion of appropriate BCG therapy invades the subepithelial connective tissue) disease. Adequate BCG therapy includes a minimum of 5 of 6 induction courses (adequate induction) and 2 of 3 maintenance courses or 2 of 6 secondary induction courses. Defined.
5. Had ≧2 cystoscopic surgeries in the most recent ≦8 weeks prior to study entry and confirmed the presence of high-risk NMIBC as defined in entry criteria #4, including complete TURBT.
6. Papillary disease completely resected at study entry, residual CIS is allowed.
7. Willingness to provide tumor tissue samples (archival or fresh biopsy, including CIS). Archived tissue must be available and sufficient for biomarker analysis. Samples must be within 6 months of screening and must include tissue representatives from each segment of the bladder suspected of having CIS disease.
8. Not eligible or elected to undergo radical cystectomy.
9. ECOG performance status 0-1.
10. Intention to avoid pregnancy based on the following criteria:
o Male childbearing participants will take appropriate precautions from Screening to 90 days after the last dose of study treatment to avoid becoming the father of a child (with at least 99% certainty) You must agree to this and refrain from donating sperm during this period. Participants should be informed and understood of an approved method that is at least 99% effective in preventing pregnancy.
o Females of childbearing potential must have a negative serum pregnancy test at Screening and take appropriate precautions (at least 99% certainty) to avoid pregnancy from Screening until 6 months after the last dose of study treatment. must agree to take Participants should be informed and understood of an approved method that is at least 99% effective in preventing pregnancy.
o Women who are infertile (ie, surgically infertile by hysterectomy and/or bilateral oophorectomy or amenorrhea for ≥12 months and at least 50 years of age) are eligible.

Investigational Treatment Information Table 7 presents the investigational treatment information for retifanlimab and epacadostat, respectively. In clinic-administered epacadostat visits, it should be administered immediately prior to initiation of retifanlimab infusion. Dose modification of retifanlimab and epacadostat is not allowed.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. The materials, methods, and examples are illustrative only and not intended to be limiting.

Claims (43)

A method of treating cancer in a patient, comprising:
(i) epacadostat or a pharmaceutically acceptable salt thereof at a dose of about 400 mg to about 700 mg on a free base basis twice daily;
(ii) an antibody that binds to human PD-1, or an antigen-binding fragment thereof, wherein said antibody comprises (ii-1) variable heavy (VH) complementarity determining regions (CDR) 1, VH CDR2, and VH CDR3; (ii-2) a VL domain comprising variable light (VL) CDR1, VL CDR2, and VL CDR3;
(a) said VH CDR1 comprises the amino acid sequence SYWMN (SEQ ID NO: 6);
(b) said VH CDR2 comprises the amino acid sequence VIHPSDSETWLDQKFKD (SEQ ID NO: 7);
(c) said VH CDR3 comprises the amino acid sequence EHYGTSPFAY (SEQ ID NO: 8);
(d) said VL CDR1 comprises the amino acid sequence RASESVDNYGMSFMNW (SEQ ID NO: 9);
(e) said VL CDR2 comprises the amino acid sequence AASNQGS (SEQ ID NO: 10);
(f) the above method, wherein said VL CDR3 comprises the amino acid sequence QQSKEVPYT (SEQ ID NO: 11); 2. The method of Claim 1, wherein said epacadostat is administered as a free base. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 400 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 425 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 450 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 475 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 500 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 525 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 550 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 575 mg twice daily. 3. The method of claim 2, wherein said epacadostat is administered at a dose of about 600 mg twice daily. 12. The method of any one of claims 1-11, wherein said antibody is administered at a dose of about 500 mg. 12. The method of any one of claims 1-11, wherein the antibody is administered at a fixed dose of about 500 mg once every four weeks. 12. The method of any one of claims 1-11, wherein the antibody is administered at a fixed dose of about 375 mg once every three weeks. 15. The method of any one of claims 1-14, wherein said antibody is administered via intravenous administration. 16. The method of any one of claims 1-15, wherein said anti-VH domain comprises the amino acid sequence shown in SEQ ID NO:4. 17. The method of any one of claims 1-16, wherein said antibody comprises a heavy chain, said heavy chain comprising the amino acid sequence shown in SEQ ID NO:2. 18. The method of any one of claims 1-17, wherein the VL domain comprises the amino acid sequence shown in SEQ ID NO:5. 19. The method of any one of claims 1-18, wherein said antibody comprises a light chain, said light chain comprising the amino acid sequence shown in SEQ ID NO:3. 20. The method of claims 1-19, wherein said VH domain comprises the amino acid sequence shown in SEQ ID NO:4 and said VL domain comprises the amino acid sequence shown in SEQ ID NO:5. 21. Any of claims 1-20, wherein said antibody comprises a heavy chain and a light chain, said heavy chain comprising said amino acid sequence shown in SEQ ID NO:2, and said light chain comprising said amino acid sequence shown in SEQ ID NO:3. or the method according to item 1. 22. The method of any one of claims 1-21, wherein said antibody comprises an Fc region of the IgG4 isotype and an IgG4 hinge domain comprising stabilizing mutations. The method of any one of claims 1-22, wherein said antibody is a humanized antibody. 24. The method of any one of claims 1-23, wherein the cancer is a solid tumor. The cancer is skin cancer, lung cancer, lymphoma, sarcoma, bladder cancer, ureteral cancer, urethral cancer, and urachal cancer, gastric cancer, cervical cancer, liver cancer, breast cancer, renal cancer, head and neck cancer, squamous cell cancer, colon cancer, endometrial cancer, anal cancer, and a tumor with microsatellite instability-high (MSI-H), mismatch repair deficient (dMMR), or DNA polymerase epsilon exonuclease domain mutation-positive disease. 25. The method according to any one of 1 to 24. The cancer is cholangiocellular carcinoma, melanoma, non-small cell lung cancer, small cell lung cancer, Hodgkin's lymphoma, urothelial carcinoma, gastric cancer, hepatocellular carcinoma, Merkel cell carcinoma, triple negative breast cancer, renal cell carcinoma, head and neck cancer The method according to any one of claims 1 to 24, which is squamous cell carcinoma and colorectal cancer. The method of any one of claims 1-24, wherein the cancer is squamous cell carcinoma of the anal canal. 25. The method of any one of claims 1-24, wherein the cancer is Merkel cell carcinoma. 25. The method of any one of claims 1-24, wherein the cancer is endometrial cancer. 25. The method of any one of claims 1-24, wherein the cancer is cervical cancer. 25. The method of any one of claims 1-24, wherein the cancer is renal cancer. 32. The method of claim 31, wherein the cancer is renal clear cell carcinoma. 25. The method of any one of claims 1-24, wherein the cancer is lung cancer. 34. The method of claim 33, wherein said cancer is adenocarcinoma of said lung. 34. The method of claim 33, wherein the cancer is squamous cell carcinoma of the lung. 34. The method of claim 33, wherein said cancer is non-small cell lung cancer. 25. The method of any one of claims 1-24, wherein the cancer is head and neck cancer. 38. The method of claim 37, wherein the cancer is head and neck squamous cell carcinoma. 25. The method of any one of claims 1-24, wherein the cancer is bladder cancer. 40. The method of claim 39, wherein said bladder cancer is high-risk BCG non-responsive non-muscle invasive bladder cancer. 41. The method of any one of claims 1-40, wherein the cancer is microsatellite stable (MSS). 41. The method of any one of claims 1-40, wherein the cancer is PD-L1 positive. 42. The method of any one of claims 1-41, wherein the cancer is microsatellite stable (MSS) and PD-L1 positive.